Tesi sul tema "Mouth Diseases Diagnosis"

History has recorded the antiquity of serious infections in the region of the head and neck. Today, our community still experiences major life-threatening infections in these anatomical locations, which pose significant management difficulties to the oral and maxillofacial surgeon. The aim of this thesis is to review the aetiology, diagnosis and treatment of some bacterial infections involving structures of the head and neck. Such infections may spread, causing serious complications with severe morbidity and occasionally death. This theses deals only with infections of bacterial origin and does not attempt to cover viral, or fungal agents or the chronic specific diseases of tuberculosis and syphilis, and makes no attempt to address the old question of focal infection. The literature review relates especially to Ludwig’s Angina which was first described so dramatically in 1836. To this day it remains as a clinically potentially lethal disease despite the progress of modern medicine. Numerous descriptions in the literature warn of the rapid appearance of symptoms and the danger of respiratory obstruction when management of the airway is not satisfactorily undertaken. Both odontogenic and non-odontogenic causes of orofacial and neck infections are reviewed. Odontogenic problems are given special emphasis as they are now of major concern. The significance of the potential fascial spaces in the face and neck which allow the spread of dental infections is also highlighter. A thorough knowledge of these anatomical relationships is still of the utmost importance to the surgeon if he is to be successful in treatment. The principle of surgical drainage of pus is as important in 1990 as it was 150 years ago. The biological basis for the onset and progress of such fulminating infections in the head and neck region is still poorly understood. One constant need is that the bacteria, both aerobic and anaerobic, be correctly identified. Microbiological techniques are constantly improving and provide an important adjuvant investigation, which then allows the surgeon to provide the most appropriate antimicrobial therapy. Principal to the many aspects of treatment is the ability to maintain the airway of the patient and to provide the depth of anaesthesia necessary to undertake the required surgery. Major bacterial orofacial infections may have severe local and far-reaching systemic effects. Such complications are discussed in all their ramifications. It should be realised that the presentation of these patients at a late stage, when complications have already supervened, may make diagnosis difficult. There is always a necessity to ensure that the underlying cause of the disease is accurately defined and that complication are not allowed to progress further. Finally, a retrospective study of the management of 90 patients with major bacterial orofacial infections who have been treated at Westmead Hospital is presented. The outcome of this study of some major bacterial orofacial infections of the head and neck is the need to stress the importance of urgent surgical management and maintenance of the airway, together with the microbiological determination of the causative organisms and their sensitivities, so that other than empirical antibiotics can be instituted early. This must be combined with an upgrading of the patients’ medical and dental status. It was demonstrated that, in the majority of these patients, ignorance and fear combined with a lack of routine dental care resulted in major infections arising from relatively simple odontogenic causes such as dental caries, periodontal disease and pericoronal infection related to impacted teeth. Without doubt, the immediate care of these patients demanded intensive management. However, it is important to recognise that dental education forms an integral part not only of the recovery programme for the afflicted patient, but also as a community health preventive measure of profound significance.

Master of Dental Surgery
History has recorded the antiquity of serious infections in the region of the head and neck. Today, our community still experiences major life-threatening infections in these anatomical locations, which pose significant management difficulties to the oral and maxillofacial surgeon. The aim of this thesis is to review the aetiology, diagnosis and treatment of some bacterial infections involving structures of the head and neck. Such infections may spread, causing serious complications with severe morbidity and occasionally death. This theses deals only with infections of bacterial origin and does not attempt to cover viral, or fungal agents or the chronic specific diseases of tuberculosis and syphilis, and makes no attempt to address the old question of focal infection. The literature review relates especially to Ludwig’s Angina which was first described so dramatically in 1836. To this day it remains as a clinically potentially lethal disease despite the progress of modern medicine. Numerous descriptions in the literature warn of the rapid appearance of symptoms and the danger of respiratory obstruction when management of the airway is not satisfactorily undertaken. Both odontogenic and non-odontogenic causes of orofacial and neck infections are reviewed. Odontogenic problems are given special emphasis as they are now of major concern. The significance of the potential fascial spaces in the face and neck which allow the spread of dental infections is also highlighter. A thorough knowledge of these anatomical relationships is still of the utmost importance to the surgeon if he is to be successful in treatment. The principle of surgical drainage of pus is as important in 1990 as it was 150 years ago. The biological basis for the onset and progress of such fulminating infections in the head and neck region is still poorly understood. One constant need is that the bacteria, both aerobic and anaerobic, be correctly identified. Microbiological techniques are constantly improving and provide an important adjuvant investigation, which then allows the surgeon to provide the most appropriate antimicrobial therapy. Principal to the many aspects of treatment is the ability to maintain the airway of the patient and to provide the depth of anaesthesia necessary to undertake the required surgery. Major bacterial orofacial infections may have severe local and far-reaching systemic effects. Such complications are discussed in all their ramifications. It should be realised that the presentation of these patients at a late stage, when complications have already supervened, may make diagnosis difficult. There is always a necessity to ensure that the underlying cause of the disease is accurately defined and that complication are not allowed to progress further. Finally, a retrospective study of the management of 90 patients with major bacterial orofacial infections who have been treated at Westmead Hospital is presented. The outcome of this study of some major bacterial orofacial infections of the head and neck is the need to stress the importance of urgent surgical management and maintenance of the airway, together with the microbiological determination of the causative organisms and their sensitivities, so that other than empirical antibiotics can be instituted early. This must be combined with an upgrading of the patients’ medical and dental status. It was demonstrated that, in the majority of these patients, ignorance and fear combined with a lack of routine dental care resulted in major infections arising from relatively simple odontogenic causes such as dental caries, periodontal disease and pericoronal infection related to impacted teeth. Without doubt, the immediate care of these patients demanded intensive management. However, it is important to recognise that dental education forms an integral part not only of the recovery programme for the afflicted patient, but also as a community health preventive measure of profound significance.

Foot-and-mouth disease (FMD) is an economically devastating picornaviral disease affecting over 40 species of cloven-hoofed animals. The virus occurs as seven immunologically distinct serotypes which are characterized by high levels of intra- and intertypic variation. The three South African Territories (SAT) serotypes 1-3 are endemic to sub-Saharan Africa, a region where the epidemiology of the disease is particularly complex due to the presence of six of the seven serotypes, the role of wildlife in virus maintenance and the apparently higher levels of variation in the endemic serotypes. These factors make it imperative to establish methods suited to elucidating the regional epidemiology. One of the integral parts of this process is the genetic characterization of regionally representative viruses in order to assess the variation in the field and to clarify the role of wildlife. Nucleotide sequence data and methods suited to studying the SAT-types are however limited. A first priority was therefore to establish a PCR-based nucleotide sequencing technique targeting the highly immunogenic and phylogenetically informative 1D genome region encoding the VP1 protein. The screening of multiple serotypes and subtypes prevalent on the African continent confirmed that this method was robust and well-suited to molecular epidemiological studies in the southern Africa region. The method was first applied in the characterization of FMD virus recovered from the reproductive tract of free-living African buffalo in the Kruger National Park. Nucleotide sequencing assisted in authentication of the results and indicated that carrier status was likely, but it was not possible to unequivocally demonstrate persistent infection of FMDV. In a separate study, the role of impala antelope (Aepyceros melampus) in the epidemiology of the disease in South Africa was assessed. Genetic characterization of impala and African buffalo (Syncerus caffer) viruses collected over an eleven year period confirmed that inter-species transmission occurred on several occasions and that virus can persist in impala populations for more than 12 months. Inter-species transmission and investigation of the possible mechanisms facilitating virus transmission from persistently infected buffalo focussed on the Kruger National Park in South Africa. In order to ensure regional relevance the study was broadened to incorporate buffalo populations throughout southern Africa. Viruses of the three SAT-types recovered from diverse African buffalo populations were therefore characterized. The results reveal that independently evolving viral lineages occur in distinct geographical regions for each of the SAT-types examined and that the levels of intratypic variation are in the order of 52 - 55 % on nucleotide level across the genome region characterized. Given the strict locality-specific grouping of buffalo viruses the likely usefulness of this database for tracing the origin and course of contemporary and historical SAT-type outbreaks was investigated. Molecular epidemiological studies conclusively show that buffalo are indeed the ultimate source of infection for susceptible cloven-hoofed animals occurring in close proximity, that interspecies transmission occurs between cattle and antelope and that trans-boundary transmission of virus remains a threat to disease security in southern African countries.
Thesis (PhD (Microbiology))--University of Pretoria, 2007.
Microbiology and Plant Pathology
unrestricted

Foram avaliados 624 prontuários clínicos, pertencentes a uma Clínica particular de Estomatologia localizada na cidade de Lins-SP, os quais foram confeccionados no período de 1989 a 2014. Procurou-se investigar a prevalência de alterações sistêmicas e de alterações bucais, buscando-se ainda determinar o índice de correlação entre estas duas alterações, em função do gênero, de grupo étnico-racial, dos fatores de risco (tabagismo e etilismo) e da faixa etária. Os dados coletados foram submetidos à análise estatística, tendo sido estabelecido um intervalo de confiança de 95% e nível de significância de 5%. Utilizou-se o teste do Qui Quadrado para avaliar a associação entre variáveis e a Regressão Logística Binária para determinar os fatores preditores das principais alterações bucais observadas nesta população deste estudo, as neoplasias, as lesões precursoras e as alterações glandulares. Foi constatado que houve uma maior procura pelos serviços estomatológicos por indivíduos do gênero feminino (63,5%), leucodermas (84,3%), não fumantes (64,9%) e não etilistas (86,7%), sem prevalência de idade. Em um total de 1.902 doenças sistêmicas houve prevalência das doenças oftálmicas (54%), alterações imunológicas (38,5%), distúrbios neurológicos (36,4%) e alterações cardiovasculares (35,1%). As doenças bucais apresentaram um total de 866 alterações patológicas, abrangendo 99,52% da amostra, com prevalência das doenças inflamatórias (27,4%), lesões traumáticas (24,0%), variações da normalidade (22,4%) e doenças glandulares (19,1%). Foram verificadas as associações entre neoplasias benignas bucais e neoplasias em geral, de neoplasias malignas bucais e doenças cardiovasculares, ou com as alterações imunológicas. Foram calculados os índices de correlação entre alterações glandulares bucais e cardiovasculares, distúrbios neurológicos, doenças infectocontagiosas, geniturinárias, ósseas, distúrbios hematológicos e neoplasias. As lesões traumáticas foram associadas com alterações cardiovasculares, doenças ósseas e doenças oftálmicas. Idade maior de 50 anos e etilismo pareceram fatores preditores para neoplasias bucais e as alterações imunológicas fatores protetores. Quanto às lesões traumáticas bucais, os fatores preditores foram idade maior que 50 anos e doenças oftálmicas. Para as lesões glandulares de boca os fatores preditivos foram alterações neurológicas, geniturinárias e hematológicas.
A total of 624 clinical records belonging to a private Clinic of Stomatology located in Lins - SP were evaluated, which were made in the period of 1989 to 2014. We sought to investigate the prevalence of systemic diseases and oral changes, seeking to determine the correlation index between these two changes, depending on gender, ethnic and racial group, the risk factors (smoking and drinking) and age group. Data were statistically analyzed, having been established a confidence interval of 95% and 5% significance level. We used the Chi square to evaluate the association between variables and Binary Logistic Regression to determine the predictors of major oral changes observed in this population study, neoplasms, precursor lesions and glandular changes. It was found that there was a greater demand for Stomatological services by female subjects (63.5%), Caucasian (84.3%), non-smokers (64.9%) and non-alcoholic (86.7%) without prevalence of age. In a total of 1,902 systemic diseases, there was prevalence of ophthalmic diseases (54%), immunological disorders (38.5%), neurological disorders (36.4%), cardiovascular disorders (35.1%). The oral diseases were a total of 866 pathological changes, including 99.52% of the sample, with a prevalence of inflammatory diseases (27,4%), traumatic injuries (24.0%), normal variation (22.4%) and glandular disease (19.1%). Associations were found between oral cancer and benign neoplasms in general, mouth cancers and cardiovascular diseases, or immune changes. The correlation ratios between oral and cardiovascular glandular changes were calculated, neurological disorders, infectious, genitourinary and bone diseases, hematological disorders and neoplasms. Traumatic injuries have been associated with cardiovascular, bone diseases and ophthalmic diseases. Age higher than 50 years and alcohol consumption seemed predictive factors for oral cancer and immunological changes being protective factors. As for the mouth traumatic injuries, the predictors were age greater than 50 years and ophthalmic diseases. For glandular lesions of the mouth the predictive factors were neurological, genitourinary and hematological changes.

A síndrome de ardência bucal (SAB) é classicamente descrita como uma doença orofacial crônica, caracterizada pela presença do sintoma de ardência ou queimação na mucosa oral clinicamente normal. A SAB possui significativa predileção pelo gênero feminino e alta prevalência em indivíduos acima dos 40 anos de idade. Sua etiologia, embora desconhecida, é considerada multifatorial e, frequentemente, associada a fatores locais, sistêmicos, psicogênicos e neuropáticos. Diversos tratamentos são propostos para a SAB, porém considerados empíricos e ineficazes. Os objetivos do presente estudo foram investigar os fatores associados ao diagnóstico da SAB e a eficácia da acupuntura como modalidade terapêutica. Os dados de 95 pacientes diagnosticados com SAB no ambulatório da Disciplina de Estomatologia Clínica da FOUSP no período de Janeiro de 1993 a Fevereiro de 2005 foram avaliados retrospectivamente. Os critérios de inclusão e exclusão considerados no diagnóstico da SAB foram estabelecidos pela queixa de ardência bucal e ausência de sinais clínicos visíveis na mucosa oral. A relação entre os fatores prognósticos (classificação, localização, intensidade, natureza e escore EVA) e os fatores associados (xerostomia, hipossalivação, pH salivar, candidose bucal, prótese dentária removível, comorbidades, medicação sistêmica, ansiedade, depressão, estresse, cancerofobia, idade, gênero, etnia, e hábitos), foi analisada estatisticamente através do teste exato de Fisher, Regressão de Poisson univariada e multivariada e o teste de Mann-Whitney. Para a análise da resposta terapêutica da acupuntura foi utilizado o teste de Wilcoxon para verificar diferenças entre a sintomatologia avaliada através de EVA (0-100), intensidade (suave-moderada-grave) e frequência (intermitente-contínua), antes e depois do tratamento acupuntura. O nível de significância de 5% (p<0,05) foi considerado. Os resultados obtidos evidenciaram significativa predileção da SAB pelo gênero feminino, presente em 87 (91,58%) mulheres, especialmente de etnia Caucasiana. A média de idade foi de 59,76 anos (32-86 anos), sendo mais prevalente na faixa etária entre 50 e 70 anos. Os Tipos 1 e 2 de SAB foram os mais prevalentes, e a língua foi o local mais afetado, referido por 79 (83,16%) pacientes. A média de duração dos sintomas foi de 3 anos (2 meses a 20 anos). O escore médio mensurado por EVA foi de 80,63 (30-100), sendo mais prevalentes os sintomas de intensidade grave e contínua. Os fatores mais frequentes foram: uso de medicações sistêmicas, constatado em 75 (78,95%) pacientes, comorbidades, em 73 (76,84%), e xerostomia, em 33 (34,74%) indivíduos. Dentre as inúmeras análises realizadas, não foi evidenciada qualquer correlação estatisticamente significativa (p>0,05) entre os fatores estudados e a SAB. Quanto à resposta terapêutica da acupuntura, evidências quanto à sua eficácia no tratamento da SAB foram constatadas pela diminuição estatisticamente significante no escore EVA (p<0,01), assim como verificado para a intensidade, que variou de grave a ausente (p<0,01), e para a frequência dos sintomas, variando de contínua a ausente ou intermitente (p<0,01), no período estudado. De acordo com os resultados obtidos no presente estudo, concluímos que fatores frequentemente associados à SAB não evidenciaram correlações significativas com esta síndrome em nossa casuística e que a acupuntura se mostrou eficaz na melhora da sintomatologia de ardência bucal referida pelos pacientes.
Burning mouth syndrome (BMS) is classically described as a chronic orofacial disease, characterized by burning mouth symptom in oral mucosa clinically normal. The BMS has a significant predilection for female gender and high prevalence in adults over 40 years old. Its etiology, although unknown, is considered multifactorial and often associated to local, systemic, psychogenic and neuropathic factors. Several treatments are proposed for BMS, although considered empirical and ineffective. The aims of this study were to investigate clinical factors associated with diagnosis of BMS and to evaluate the effectiveness of acupuncture as a therapeutic modality. Data from 95 patients diagnosed with BMS the outpatient clinic of the Department of Stomatology, FOUSP, between January, 1993 and February, 2005 were assessed retrospectively. The inclusion and exclusion criteria considered in the diagnosis of BMS were established by complaints of burning mouth and no clinical signs visible in the oral mucosa. The correlation between prognostic factors (classification, location, intensity, nature and VAS score) and the associated factors (xerostomia, hyposalivation, salivary pH, oral candidiasis, removable dental prosthesis, comorbidities, systemic medication, anxiety, depression, stress, cancerophobia, age, gender, ethnicity, and habits), was statistically evaluated by means of Fisher exact test, Poisson regression univariate and multivariate analysis and the Mann-Whitney test. To investigate the therapeutic response of acupuncture, the Wilcoxon test was used to identify differences between symptoms assessed by VAS (0-100), intensity (mild-moderate-severe) and frequency (continuous-intermittent) before and after acupuncture treatment. The significance level of 5% (p<0.05) was considered. The results showed a significant preference of BMS in gender female, present in 87 (91.58%) women, especially Caucasian ethnicity. The mean age was 59.76 years (32-86 years), most prevalent between the ages of 50 and 70 years. Types 1 and 2 of BMS were the most prevalent, and the tongue was the most affected site, reported by 79 (83.16%) patients. The mean duration of symptoms was 3 years, ranging from 2 months to 20 years. Regarding symptom intensity, the mean score measured by VAS was 80.63 (30-100), and the most frequent symptoms were severe and continuous. Among of considered factors, the most frequent were: use of systemic medications found in 75 (78.95%) patients, comorbidities conditions, in 73 (76.84%), and xerostomia, in 33 (34.74%) subjects. Among the numerous analysis carried out, no statistically significant correlation was evidenced (p>0.05) between all the considered factors and BMS. Regarding the therapeutic response of acupuncture, considerable evidences concerning its efficacy in the treatment of BMS were observed by a statistically significant decrease in VAS score (p<0.01), as observed for the intensity, which ranged from severe to absent (p<0.01), and the frequency of symptoms, ranging from continuous to intermittent or absent (p<0.01) during the study period. According to the results obtained in this study, we conclude that factors often associated with BMS did not show statistically significant correlations with this syndrome in our series, and that acupuncture was effective in reducing the symptoms intensity of burning mouth referred by patients.

Epiteloidcellig granulomatos (ECG) är en patologanatomisk diagnos (PAD) av en inflammationstyp som kan ses i orala vävnadsbiopsier. Inflammationsbilden visar infiltrat av inflammationsceller och epiteloidcelliga granulom med eller utan jätteceller, därtill kan ödematös vävnad och fibros presenteras. Makroskopiskt ses en symtombild med t ex läpp- och ansiktssvullnad, munvinkelragader och gingivala hyperplasier. Klinisk ställs exklusionsdiagnosen orofacial granulomatos (OFG). Etiologin är multifaktoriell och kan ses vid exempelvis Crohns sjukdom (CS) och sarkoidos. På Avdelning för Oral Patologi, Malmö Tandvårdshögskola, sker PAD av biopsier som vid ECG diagnoskodas med 995, vilket tilldelats 152 remissfall. Metoden utgörs av en retrospektiv uppföljning av dessa remisser med syfte att undersöka etiologin, utfört remitteringshandhavande, samt för att få en uppfattning om den orala biopsin bidragit till eventuell systemisk sjukdomsdiagnos och huruvida återkoppling mellan remissinstanserna förekommit. Vidare undersöktes om ett eventuellt samarbete mellan tandläkare och läkare praktiserats.Antalet deltagande vårdgivare i studien visar på ett intresse för ECG och dess etiologi. Misstänkt etiologi var i de flesta fall CS vilket stämde överens med ställd diagnos. Remitteringshandhavandet visade i vissa fall på bristande återkoppling mellan inkluderade vårdinstanser och vårdgivare samt avsaknad av förutbestämda remitteringsvägar. Andra remissfall visade väl fungerande samarbete mellan vårdyrken och instanser samt att den orala biopsin i vissa fall bidragit till ställd systemisk sjukdomsdiagnos. Författarna önskar att studien i förlängningen ska kunna bidra till väl fungerande kommunikation mellan berörda vårdgivare med förhoppning om snabbare handhavande, säkrare patienthantering med tidigare ställd diagnos och minskat lidande för patienterna.
Epithelioid cell granulomatosis (ECG) is a diagnosis of anatomical pathology (PAD) and a type of inflammation seen in oral biopsies. ECG presents with histological features such as epithelioid cell granuloma with or without presence of multinucleated giant cells, inflammatory cells and occasionally oedematous fibrotic changes. Clinical symptoms presents as e.g. enlargement of lips, facial swelling, angular cheilitis and gingival hyperplasia. Orofacial granulomatosis (OFG) is the clinical diagnosis given PAD ECG with a multifactorial aetiology e.g. Crohn’s disease (CS) and sarcoidosis. The Department of Oral Pathology, Faculty of Odontology, Malmö University, determine PAD of oral biopsies that when given ECG are marked with the code of diagnosis 995, given 152 referrals. This retrospective follow-up of referrals with PAD ECG aims to investigate represented aetiology, management of referrals and to get a notion of whether the oral biopsy has contributed to systemic diagnosis, if there has been any feedback between included instances and collaboration between dentists and physicians. Number of included participants could be interpreted as an interest in ECG, which most frequently showed to be CS, agreeing with later confirmed diagnosis. The management in some cases of referral lacked feedback between health professionals as well as absence of predetermined ways of referral. Others showed cooperation and that the oral biopsy had contributed to diagnosis of systemic disease. The authors wish that this study will contribute to a well functional communication between health professionals, anticipating a more efficient and secure management of patients, improvement of early diagnosis and decreased patient discomfort.

Thesis (MSc)--Stellenbosch University, 2002.
ENGLISH ABSTRACT: The aim of this study at the Brooklyn Chest Hospital (BCH) was to assess the change in the oral carriage of Candida species in twenty-nine patients co-infected with the Human Immunodeficiency Virus (HIV) and Tuberculosis (TB), before and after anti-fungal treatment. Each patient accepted onto the study underwent a comprehensive oral and peri-oral examination where the presence, site and clinical features of all oral and peri-oral lesions were recorded. The purpose of the examination was to provide a clinical diagnosis of oral candidasis. Each patient was also asked to provide a sample of oral fluid for laboratory analysis. This was collected using an oral rinse. The results of a variety of laboratory investigations were used to identify the species of Candida obtained from the oral rinse. Both the oral and peri-oral examination and the oral rinse procedure were repeated after one month and at three months. A sample from each oral rinse was inoculated on CHROMagar Candida chromogenic medium (CHROMagar Candida, France, Paris). CHROMagar is used for the isolation and presumptive identification of Candida sp. from other yeasts on the basis of strongly contrasted colony colours, which are produced by the reactions of species-specific enzymes with a proprietary chromogenic substrate. After forty-eight hours the CHROMagar plate was examined for growth, when a record of colony morphology and colour was made. A single sample from each different colour-coded colony was taken and streaked onto a Sabouraud plate (Oxoid, Basingstake, England) and then incubated for forty eight hours at thirty-seven degrees centigrade. A variety of laboratory investigations were subsequently carried out on a single colony taken from the Sabouraud agar plate (Oxoid). The results of these tests were used to identify the individual species of Candida isolated from each oral rinse. Oral candidasis was the most prevalent oral lesion observed on admission and at three months. Six different species of Candida were identified during this study, namely Candida albicans, Candida dubliniensis, Candida krusei, Candida glabrata, Candida parapsilosis, and Candida tropicalis. C.albicans was the most commonly identified species in study population. Candida dubliniensis was isolated and identified for the first time in a South African HIV population. Each specimen of Candida sp. identified by laboratory analysis was tested for sensitivity to Nystatin, Amphotericin B and Fluconazole anti-fungal agents. An additional sensitivity test was performed using Ajoene and Allicin (extracts of garlic) to assess the comparative antifungal properties of these compounds.
AFRIKAANSE OPSOMMING: Die doelwit van hierdie studie by die Brooklyn Borshospitaal (BCH) was om die verandering in orale draerstatus van die Kandida spesies in nege-en-twintig HIVfTB koïnfekteerde pasiënte vas te stel, voor- en na antifungale behandeling. Elke pasiënt in die studie het 'n volledige intra- en ekstra-orale ondersoek ondergaan. Die teenwoordigheid, area en kliniese voorkoms van alle letsels is noteer. Die doel van die ondersoek was om 'n kliniese diagnose van orale kandidiase te verkry. 'n Monster orale vloeistof is geneem van elke pasiënt vir laboratorium analise. Die monster is in die vorm van 'n mondspoel geneem. Verskeie toetse is gedoen om die verskillende Kandida spesies in elke monster te identifiseer. Die orale- en ekstra-orale ondersoek sowel as die mondspoelmonster is na 1 en 3 maande herhaal. Elke mondspoelmonster is op CHROMagar Kandida chromogene medium (CHROMagar Candida, France) inokuleer. CHROMagar word gebruik vir die vermoedelike identifikasie en isolasie van Kandida spesies teenoor ander swamme. Dit word gedoen op die basis van kontrasterende koloniekleure, wat teweeggebring word deur spesie-spesifieke ensiemreaksies op 'n chromogene substraat. Die CHROMagar plate is na 48 uur ondersoek vir groei en die kolonie-morfologie en - kleur is noteer. 'n Enkel monster. is geneem van elke verskillende kolonie (geskei op kleur) en is uitgestreep op 'n Saboraud plaat (Oxoid, Basingstoke, England). Dit is dan vir 48 uur inkubeer teen 37°C. Verskeie laboratorium ondersoeke is daarna uitgevoer op 'n enkel kolonie geneem vanaf die Saboraud agar plaat (oxoid). Die resultate van die ondersoeke is gebruik om individuele spesies van Kandida te identifiseer. Orale Kandidiase was die mees algemene orale letsel geïdentifiseer by toelating en 3 maande ondersoeke. Ses verskillende spesies Kandida is identifiseer tydens die studie, naamlik: Kandida albicans, K.dubliniensis, K.Krusei, K.glabrata, K.parapsilosis en K.tropicalis. K.albicans was die mees algemeen identifiseerde spesie in die studiepopulasie. K.dubliniensis is vir die eerste keer in Suid-Afrika in 'n HIV<+lpopulasie isoleer en geïdentifiseer. Elke monster van identifiseerde Kandida spesies is getoets vir sensitiwiteit teenoor Nistatien, Amfotensien B en Flukonasool. Addisioneel is ook getoets vir sensitiwiteit teenoor Ajoene en Allicin (knoffelekstrakte).

The Foot-and-mouth disease virus (FMDV) affects cloven-hoofed animals and is endemic in most parts of Africa, South America and southern Asia. South Africa is considered a FMDV-free zone but the virus is maintained within the wildlife in the Kruger National Park (KNP), making mitigation of outbreaks a high priority. Diagnostic methods are usually costly due to the high production cost of the reagents used, meaning that regular monitoring and diagnosis of animals around the KNP for FMDV is expensive due to the large amounts of serum continuously being tested. I propose an alternative plant expression platform for the local production of more cost effective diagnostic reagents capable of distinguishing between infected and vaccinated animals (DIVA). I selected the non-structural 3ABC polyprotein of FMDV to express, as it is a suitable candidate as a coating antigen in a competitive enzyme linked immunosorbent assay (C-ELISA) for the detection of neutralizing antibodies in livestock sera. I also chose other variations of the full polyprotein (3AB, 3AB1 and 3B) for expression as they have previously been shown to be effective in FMDV diagnosis. I also selected a second reagent to be expressed: this was the CRAb-FM27 single chain variable fragment (scFv), which binds a 3B epitope on the 3ABC polyprotein and has previously shown to be effective as a competing antibody in a C-ELISA. The 3B antigen and the scFv were successfully expressed and purified from N. benthamiana, which to my knowledge is the first time either has been shown. The plant produced scFv successfully bound the 3B antigen in an I-ELISA. Separately, the plant produced 3B antigen could be used to successfully differentiate FMDV infected and vaccinated guinea pig serum in an I-ELISA. However, testing of these reagents in tandem within a C-ELISA to DIVA sera was inconclusive, and further research is required to optimise C-ELISA conditions.

The requirements for prompt diagnosis of foot-and-mouth disease (FMD) during outbreaks, and the need to establish robust laboratory testing capacity within FMD-endemic countries, have motivated the development of point-of-care tests (POCTs) to support current diagnostic strategies. Despite numerous publications detailing the design of platforms and assays for this purpose, the majority have only been evaluated in laboratory settings, using protocols incompatible for use in challenging environments. To address this gap, this thesis describes the development of an end-to-end molecular toolbox for the detection and characterisation of FMD virus (FMDV) RNA in decentralised settings. A critical review and multiway comparison of seven assay formats and 11 sample preparation methods revealed that reverse transcription loop-mediated isothermal amplification (RT-LAMP) and real-time reverse transcription PCR (rRT-PCR) POCT-formats exhibited comparable analytical and diagnostic sensitivity to their laboratory-based equivalents. Additionally, reagent lyophilisation provided a solution for cold chain and storage considerations, whilst not compromising assay performance. Both assays were compatible with simple sample preparation methods, removing the requirement for nucleic acid extraction. For example, dilution of samples in nuclease-free water enabled FMDV RNA to be detected in multiple sample types (epithelial tissue suspensions, serum, oesophageal-pharyngeal fluid and lesion swabs), from as early as one day post infection. Notably, when the robust field-ready protocols were deployed into challenging low-resource laboratory and field-settings within East Africa, POCT results (rRT-PCR = 144; RT-LAMP = 145) were consistent with clinical observations and a reference rRT-PCR, with FMDV detected from acutely infected as well as convalescent cattle. Furthermore, transitioning of East Africa-specific FMDV-typing rRT-PCR assays (for serotypes O, A, Southern African Territories [SAT] 1 and SAT 2) into a multiplex POCT-format enabled rapid identification of FMDV serotype in situ, confirming active outbreaks of both O and A. This thesis also describes the development of GoPrime, a novel real-time PCR (rPCR) primer/probe validation tool. By parameterising GoPrime with experimental data, collected to investigate the effects of primer/probe-template mismatches on cycle threshold and limit of detection, it was possible to quantitatively predict the performance of rPCR assays in silico. The work of this thesis supports the deployment of molecular POCTs into non-specialised, resource-limited and challenging settings for simple, highly sensitive and rapid detection and/or characterisation of FMDV.

This study explored and described the strategies used by nurses to manage newly diagnosed HIV positive pregnant women who do not return to the clinic within a month following diagnosis, for continuity of care. The main aim was to prevent mother-to-child transmission of HIV (PMTCT). Method: The study sites were two accredited antiretroviral- ante-natal care (ARV-ANC) clinics at the Dimbaza community health centre (CHC) and the East London Hospital Complex (ELHC) which comprises of Cecilia Makiwane and Frere hospitals, in the Buffalo City Metropolitan Municipality (BCM). The majority of health personnel at these clinics are professional nurses. An in-depth semi structured interview guide was used to collect data through focus group interviews from professional nurses who work in these units. They were required to share their experiences about intervention strategies used for newly diagnosed HIV- positive, pregnant women who fail to return for continuity of care within a month after diagnosis. Results: Follow up of these women is done by tracking them (by calling them; calling the clinic nearest to their homes and doing home visits). Decentralisation of further management and care to the nearest clinic was also cited, especially for the patients who stay far from these accredited sites. Family support was also mentioned as a strategy to intervene for the non-compliant patients. Challenges: The challenges that were encountered with these interventions include wrong contact details, wrong addresses and being evasive when visited at home. Another challenge cited was related to the stigma attached to the diagnosis and the tracking devices used, for example, the car as it is familiar to the community it serves. Despite known benefits for early initiation of HIV treatment newly diagnosed HIV- positive, pregnant women continue to refrain from accessing care after diagnosis, thus posing a risk to the transmission of HIV to the baby and further comprising their own health. They miss out on general HIV management and ante-natal care. Conclusion: The identified intervention strategies used by nurses to follow up newly diagnosed HIV- positive, pregnant women need to be reinforced and strategies put in place to control the related challenges for a better response by the patients.

"8th July 2002." Includes bibliographical references (leaves 276-308) Examines the systematics (taxonomy, phylogeny, and epiemiology) of the genus Candida using a combination of traditional and contemporary methodologies. Assesses these methods to determine their diagnostic potential to unequivocally identify and characterise species and strains of this medically and dentally important yeast genus.

"8th July 2002."
Includes bibliographical references (leaves 276-308)
vi, 308, [57] leaves : charts ; 30 cm.
Title page, contents and abstract only. The complete thesis in print form is available from the University Library.
Examines the systematics (taxonomy, phylogeny, and epiemiology) of the genus Candida using a combination of traditional and contemporary methodologies. Assesses these methods to determine their diagnostic potential to unequivocally identify and characterise species and strains of this medically and dentally important yeast genus.
Thesis (Ph.D.)--University of Adelaide, Dept. of Dentistry, 2002

碩士
國立中興大學
獸醫學系暨研究所
94
Abstract Foot-and-mouth disease (FMD) outbreaks usually result in huge economic losses. FMD is a severe, clinically acute, vesicular disease of cloven-hoofed animals including domesticated ruminants and swine, FMDV belongs to the Picornaviridae family, and is the only member of the genus Aphthovirus, Serologically, FMDV can be classified into 7 antigenically distinct serotypes, O, A, C, SAT1, SAT2, and SAT3, and Asia 1. The virus has a predilection to replicate in epithelial cells including those lining the distal oropharynx and the dorsal surface of the soft palate. In these sites, infectious virus has been found for several months or even years in a proportion of recovered ruminants but not pigs. Detection of FMDV is important for monitoring vaccination status and detection of the carries. Therefore, it is important to develop diagnostic methods for the detection of FMDV antibody. In this study, FMDV VP1 and VP2 gene of foot-and-mouth disease virus were amplified by RT-PCR. The DNA containing FMDV VP1 or VP2 gene sequences was further cloned into a prokaryotic expression vetor pET21d, respectively, named pET21d FMDV-VP1 or pET21d FMDV-VP2, respectively. The recombinant FMDV VP1 or VP2 protein was expressed by the host E. coli BL21(DE3). The protein with expected size about 25.8 kDa or 26.4 kDa was obviously observed in SDS-PAGE. The recombinant FMDV VP1 and VP2 proteins were identified by western blot assay with FMDV-infected swine serum. Expressed FMDV VP1 and VP2 proteins were purified by passing through the Ni-NTA Superflow column. Using the purified proteins as a coating antigen, FMDV-infected swine serum and vaccinated swine serum were used for ELISA test. Our results demonstrated that the recombinant FMDV VP1 protein has a better specificity than that of the VP2 recombinant protein.

博士
國立臺灣大學
獸醫學研究所
102
Foot-and-mouth disease (FMD), swine vesicular disease (SVD), vesicular stomatitis (VS), and vesicular exanthema (VE) are highly contagious vesicular diseases of animal but are not able to be differentiated clinically. For the purpose of instant detecting FMD and differentiating it from the other vesicular diseases, many methods have been developed and evaluated in recent years. The structural protein VP1 gene of FMDV O/TW/1997 and the non-structural protein 3ABC gene of FMDV O/TW/1999 were constructed, respectively, into expression vectors, which were based on an Escherichia coli expression system. Subsequently, monoclonal antibodies were generated by immunizing mice with the recombinant proteins, and they were employed to develop the complex interface measurement platforms for FMDV tests. Plate and microsphere formats were developed and evaluated for their abilities in antibody detection from serum samples against structural protein (SP-VP1) and non-structural protein (NSP-3ABC) of the FMDV. In those studies, the sandwich enzyme-linked immunosorbent assay (ELISA), singleplex Luminex and multipex Luminex (xMAP) were developed for rapid detection of the FMD antibodies. In the first study, the developed sandwich ELISA demonstrated a diagnostic sensitivity (Dsn) of 98.4 % and a diagnostic specificity (Dsp) of 100 % for naive and vaccinated pigs; the detection ability of the assay was comparable with those of PrioCHECK and UBI kits. There were 97.5, 93.4 and 66.6 % agreement between the results obtained from our sandwich ELISA and those obtained from the PrioCHECK, UBI and CHEKIT kits, respectively. The kappa statistics between our ELISA and the kits were 0.95, 0.87 and 0.37, respectively. Moreover, antibodies to nonstructural proteins of the serotypes A, C, Asia 1, SAT 1, SAT 2 and SAT 3 were also detected in sera of infected cattle. In the secondary study, sera from 64 infected, 307 vaccinated, and 280 naive pigs were tested for the FMDV-NSP antibody by the sigleplex Luminex assay. The Dsn of the assay was 100%. The Dsp of the assay was 98.7% in vaccinated pigs and 97.5% to 100% in naive pigs. Agreement between the results from the singleplex Luminex and those from a 3ABC polypeptide blocking ELISA was 96.3%, and kappa statistics gave a value of 0.92. The singleplex Luminex can detect the immune response to NSP-3ABC in swine as early as eight days post-infection as same as sandwich ELISA. Moreover, the NSP-3ABC antibody in all of the 15 vaccinated but unprotected pigs which presented vesicular lesions were detected by the singleplex Luminex assay, and the antibodies in 11 of the 15 pigs were detected this antibody by the sandwich ELISA. In the third study, an xMAP was optimized to detected antibodies to SP-VP1 and NSP-3ABC of the FMD virus simultaneously in a single serum sample. To detect SP antibodies in 661 sera from infected, naive pigs and vaccinated pigs, the DSn and DSp of the xMAP were 90.0-98.7% and 93.0-96.5%, respectively. To detect NSP antibodies, the DSn was 90% and the DSp ranged from 93.3% to 99.1%. The xMAP can detect the immune response to SP and NSP as early as 4 and 8 dpi, respectively, in the experimentally infected pigs Moreover, the SP and NSP antibodies in 15 vaccinated but unprotected pigs were detected by the xMAP. Comparing the abilities in detecting the SP and NSP antibodies in the sera of infected samples, the results from the xMAP had high positive correlation with those from the virus neutralization test (VNT) and commercially available 3ABC polypeptide blocking ELISA. However, the results of xMAP had no quantitative relationship with those of the VNT. Furthermore, the specificity was 93.3-94.9% with the blocking ELISA for detecting the FMDV-NSP antibody. These studies showed that the sensitivity and specificity of the methods developed are higher than 90%, which can be as references for diagnosis and assessment of the immune status. Furthermore, the specificities of these assays were also highlighted by the absence of cross-reactions generated by antibodies against the SVDV and VSV at different titers.