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Early diagnosis and genotyping of high-risk human papillomavirus (HR-HPV) in cervical tissue specimens is significant for cervical cancer prevention. A sensitive microplate fluorometric hybridization assay (MFHA) was designed for the detection of HPV DNA 16 and 18 in cervical tissue. Following optimization and validation of the method, 60 formalin-fixed and paraffin-embedded cervical samples representing different cervical intraepithelial neoplasia grades of HPV-associated lesions were tested to determine the sensitivity and specificity of the assay. Using consensus GP5+/6+ biotin-labeled primers to amplify a conserved region within the L1 gene, the amplicons were added to the microplate wells coated with specific probes for the hybridization of HPV 16 and 18 individually. Final detection was performed with streptavidin-AlexaFluor488 conjugated. The results were then compared with type-specific nested polymerase chain reaction (PCR) and colorimetric microplate assay. While the agreement between the results obtained by the type-specific nested PCR and fluorometric assay for the detection of both HR-HPV types was 100%, this agreement for the detection of HPV type 16 and 18 using microplate colorimetric assay was 94.2% and 85% respectively. Overall, the results of the fluorometric and colorimetric assays are promising for detecting both HR-HPV types in a large number of cervical tissue samples with the higher MFHA assay sensitivity.

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BACKGROUND: Spontaneous bacterial peritonitis is defined as an ascetic fluid infection without an evident intra-abdominal surgically treatable source. Spontaneous bacterial peritonitis is one of the severe complications in patients with cirrhosis and ascites. Without early antibiotic treatment, this complication is associated with high mortality rate; therefore, early diagnosis and treatment of spontaneous bacterial peritonitis is necessary for survival. Leukocyte esterase reagent can rapidly diagnose the spontaneous bacterial peritonitis. OBJECTIVE: This study aimed to find out the diagnostic accuracy of leukocyte esterase dipstick test for the diagnosis of spontaneous bacterial peritonitis. METHODS: A single centered hospital-based cross-sectional study was conducted during July 2013 to August 2014 on children with cirrhotic liver disease and ascites who were admitted in the Department of Pediatric Gastroenterology in Nemazee Hospital affiliated to Shiraz University of Medical Sciences (Iran). All patients underwent abdominal paracentesis, and the ascitic fluid was processed for cell count, leukocyte esterase reagent strip test (Combiscreen SL10) and culture. Spontaneous bacterial peritonitis was defined as having a polymorphonuclear count (PMN ≥250/m3) in ascitic fluid. Sensitivity, specificity, positive predictive value and negative predictive value of leukocyte esterase test were calculated according to the formula. RESULTS: Totally, 150 ascitic fluid sample of cirrhotic male patients (53.2%) and their mean age (4.33±1.88 years) were analyzed. Biliary atresia (n=44, 29.4%) and idiopathic neonatal hepatitis (n=29, 19.3%) were the most frequent etiology of cirrhosis. Also, abdominal pain (68.6%) and distension (64%) were the most common presenting complaint. Of all cases, 41 patients (27.35%) were diagnosed to have spontaneous bacterial peritonitis (PMN ≥250/mm3). Sensitivity and specificity of leukocyte esterase reagent test according to PMNs ≥250 mm3 were 87.80% and 91.74%, also on ascitic fluid culture results were 88.23% and 77.44%. Positive predictive value and negative predictive value of this test in PMNs ≥250 mm3 were 80% and 95.23% and in cases with positive culture 33.33% and 98.09% were obtained, respectively. Efficiency of leukocyte esterase reagent test in diagnosing spontaneous bacterial peritonitis, according to PMNs ≥250 mm3 and culture results were 90.66% and 78.66%. CONCLUSION: The leukocyte esterase strip test may be used as rapid test for diagnosis of spontaneous bacterial peritonitis due to its high diagnostic validity.

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BackgroundSpontaneous bacterial peritonitis is defined as an ascetic fluid infection without an evident intra-abdominal surgically treatable source. Spontaneous bacterial peritonitis is one of the severe complications in patients with cirrhosis and ascites. Without early antibiotic treatment, this complication is associated with high mortality rate; therefore, early diagnosis and treatment of spontaneous bacterial peritonitis is necessary for survival. Leukocyte esterase reagent can rapidly diagnose the spontaneous bacterial peritonitis.ObjectiveThis study aimed to find out the diagnostic accuracy of leukocyte esterase dipstick test for the diagnosis of spontaneous bacterial peritonitis.MethodsA single centered hospital-based cross-sectional study was conducted during July 2013 to August 2014 on children with cirrhotic liver disease and ascites who were admitted in the Department of Pediatric Gastroenterology in Nemazee Hospital affiliated to Shiraz University of Medical Sciences (Iran). All patients underwent abdominal paracentesis, and the ascitic fluid was processed for cell count, leukocyte esterase reagent strip test (Combiscreen SL10) and culture. Spontaneous bacterial peritonitis was defined as having a polymorphonuclear count (PMN ≥250/m3) in ascitic fluid. Sensitivity, specificity, positive predictive value and negative predictive value of leukocyte esterase test were calculated according to the formula.ResultsTotally, 150 ascitic fluid sample of cirrhotic male patients (53.2%) and their mean age (4.33±1.88 years) were analyzed. Biliary atresia (n=44, 29.4%) and idiopathic neonatal hepatitis (n=29, 19.3%) were the most frequent etiology of cirrhosis. Also, abdominal pain (68.6%) and distension (64%) were the most common presenting complaint. Of all cases, 41patients (27.35%) were diagnosed to have spontaneous bacterial peritonitis (PMN ≥250/mm3). Sensitivity and specificity of leukocyte esterase reagent test according to PMNs ≥250mm3 were 87.80% and 91.74%, also on ascitic fluid culture results were 88.23% and 77.44%. Positive predictive value and negative predictive value of this test in PMNs ≥250mm3 were 80% and 95.23% and in cases with positive culture 33.33% and 98.09% were obtained, respectively. Efficiency of leukocyte esterase reagent test in diagnosing spontaneous bacterial peritonitis, according to PMNs ≥250mm3 and culture results were 90.66% and 78.66%.ConclusionThe leukocyte esterase strip test may be used as rapid test for diagnosis of spontaneous bacterial peritonitis due to its high diagnostic validity.

ContextoA peritonite bacteriana espontânea é definida como uma infecção do fluido ascítico sem evidente origem intra-abdominal cirurgicamente tratável. A peritonite bacteriana espontânea é uma das complicações graves em pacientes com cirrose e ascite. Sem tratamento antibiótico precoce, esta complicação é associada com alta taxa de mortalidade. Portanto, o diagnóstico precoce e tratamento de peritonite bacteriana espontânea são necessários para a sobrevivência. O reagente de esterase de leucócitos pode rapidamente diagnosticar a peritonite bacteriana espontânea.ObjetivoEste estudo teve como objetivo descobrir a acurácia diagnóstica do teste com tiras de esterase de leucócitos para o diagnóstico de peritonite bacteriana espontânea.MétodosUm estudo transversal hospitalar unicêntrico foi realizado entre julho de 2013 e agosto de 2014 em crianças com cirrose hepática e ascite que foram admitidas no Departamento de Gastroenterologia Pediátrica no Hospital de Nemazee afiliado à Universidade de Ciencias Médicas de Shiraz (Irã). Todos os pacientes foram submetidos a paracentese abdominal, e o líquido ascítico foi processado para contagem de células, teste de tira de reagente de esterase de leucócitos (Combiscreen SL10) e cultura. Peritonite bacteriana espontânea foi definida como tendo uma contagem de polimorfonucleares (PMN ≥250/m3) no líquido ascítico. Sensibilidade, especificidade, valor preditivo positivo negativo do teste de esterase de leucócitos foram calculados de acordo com a fórmula.ResultadosForam analisados um total de 150 amostras de líquido ascítico de pacientes cirróticos; (53,2%) eram do sexo masculino e sua média de idade (4,33±1,88 anos). A atresia biliar (n=44, 29,4%) e hepatite neonatal idiopática (n=29, 19,3%) foram as etiologias mais frequentes de cirrose. Além disso, dor abdominal (68,6%) e distensão (64%) foram as queixas mais comuns de apresentação. De todos os casos, 41 (27,35%) foram diagnosticados com peritonite bacteriana espontânea (PMN ≥250/mm3). A sensibilidade e especificidade do teste de reagente de esterase de leucócitos segundo PMN ≥250mm3 foi de 87,80% e 91,74% e, para os resultados de cultura de líquido ascítico, de 88,23% e 77,44%. Valor preditivo positivo e valor preditivo negativo do teste em PMN ≥250mm3 foi de 80% e 95,23% e em casos com cultura positiva 33,33% e 98,09%, respectivamente. A eficiência do teste de reagente esterase de leucócitos no diagnóstico de peritonite bacteriana espontânea, de acordo com resultados de ≥250mm3 e cultura PMN, foi de 90,66% e 78,66%.ConclusãoO teste de tiras de esterase de leucócitos pode ser usado como um teste rápido para diagnóstico de peritonite bacteriana espontânea, devido a sua alta validade diagnóstica.

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A 4 year old female Holstein cow with multiple skin lesions was referred for clinical examination. Grossly, approximately 60 discrete cutaneous nodules and masses with variable sizes ranging from 1 to 5 cm were observed on both sides of flunks, shoulders, neck, back, and mammary glands. Histopathologically, the masses were composed of non-encapsulated accumulation of neoplastic round cells contained moderate to abundant amounts of finely eosinophilic granules in the cytoplasm. Cellular or nuclear pleomorphism and mitotic figures were not remarkable. Metachromasia was seen in cytoplasmic granules stained with toluidine blue. In the immunohistochemical staining, the neoplastic cells were positive for vimentin and c-kit. No immunoreactivity was seen for cytokeratin and HMB - 45. Based on these findings, multiple cutaneous mast cell tumor was diagnosed. Immunohistochemical features of mast cell tumor have not been reported previously in cattle.

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A 4 year old female Holstein cow with multiple skin lesions was referred for clinical examination. Grossly, approximately 60 discrete cutaneous nodules and masses with variable sizes ranging from 1 to 5 cm were observed on both sides of flunks, shoulders, neck, back, and mammary glands. Histopathologically, the masses were composed of non-encapsulated accumulation of neoplastic round cells contained moderate to abundant amounts of finely eosinophilic granules in the cytoplasm. Cellular or nuclear pleomorphism and mitotic figures were not remarkable. Metachromasia was seen in cytoplasmic granules stained with toluidine blue. In the immunohistochemical staining, the neoplastic cells were positive for vimentin and c-kit. No immunoreactivity was seen for cytokeratin and HMB - 45. Based on these findings, multiple cutaneous mast cell tumor was diagnosed. Immunohistochemical features of mast cell tumor have not been reported previously in cattle.(AU)

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