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Objective@#To evaluate the using value of FibroTouch and six serological models in detecting the degree of liver fibrosis in patients with chronic hepatitis B, in an attempt to provide reference for accurate diagnosis.@*Methods@#Two hundred and fifty-eight cases with chronic hepatitis B admitted to Xixi Hospital of Hangzhou from September 1, 2015 to September 1, 2017 were selected. All patients underwent liver histopathological examination and FibroTouch measurement to determine liver stiffness (LSM). Serum biochemical parameters were detected and the scoring values of six serological models were calculated. SAS 9.4 statistical software was used for statistical analysis, and the correlation between FibroTouch and the six serological models was analyzed by Spearman correlation. The diagnostic value of FibroTouch and six serological models was analyzed by receiver operating characteristic curve (ROC) based on liver histopathological findings.@*Results@#The median LSM of 258 cases with chronic hepatitis B was 9.4 (6.5-13.8) kPa. In the six serological models, the median value of aspartate transaminase to platelet ratio index (APRI), FIB-4 index, S-index, Forn’s index, PRPindex, and FIB-5 were 0.42 (0.28-0.62), 1.27 (0.78-2.03), 0.11 (0.07-0.20), 6.95 (5.89-8.51), 0.000 8 (0.000 6-0.000 9),and 38.59 (36.28-40.97). FibroTouch had positive correlation with APRI, FIB-4, S-index, Forn’s index, PRP, fibrosis stage (r= 0.73,P< 0.001) and inflammation grade, and had negative correlation with FIB-5, and both had statistical significance. The area under curve (AUC) of FT-LSM at S≥2, S≥3, S = 4 were 0.89, 0.90 and 0.85, respectively, which was significantly higher than serological models (P< 0.001). The AUC of S-index model at S≥2, S≥3, S = 4 were higher than other five serological models.@*Conclusion@#The diagnostic performance of FibroTouch is significantly better than serological model. S-index model has the best diagnostic performance in the six serological models, and the combination of S-index and FT-LSM may better diagnose the grading of liver fibrosis, and thus can be applied and promoted in clinic.
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Objective To establish a rapid and high sensitive assay of real-time fluorescence loop-mediated isothermal amplification assay for clinical detection of HIV-1 DNA.Methods Four loop primers were constructed for loop-mediated isothermal amplification ( LAMP) assay based on six conserved regions selected from HIV gene sequence .SYBR Green Ⅰdye was added into the established LAMP assay and its specificity and sensitivity were evaluated .Results The real-time fluorescence LAMP assay for the detection of HIV-1 DNA was successfully established .It was used for the detection of HIV-1 DNA among 200 patients with HIV infection, of which 195 cases were positive.Moreover, 50 healthy subjects were found HIV-1 DNA negative tested by the real-time fluorescence LAMP assay .Quantitative testing for HIV-1 DNA showed that the lowest and the highest detectable amount were 51 copies/ml and 8.21×106 copies/ml respectively, and the average amount was 5.78×105 copies/ml.HIV viral loads ranging from 1×105 to 10×105 were detected in 162 of 200 patients (83.08%).The ten times dilution method showed that the lowest detection limit of the assay was 50 copies/ml.The crossover experiment indicated that the specificity of the assay was 100%as none of HBV-DNA, HSV-DNA and HCV-RNA was determined by the assay .Conclusion The present study shows that 97.5%of the patients with HIV infection are confirmed HIV-1 DNA positive by the real-time fluorescence LAMP assay , suggesting that the real-time fluorescence LAMP assay is a rapid and sensi-tive assay with high specificity and could be applied for clinical detection of HIV-1 DNA.
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Objective To develop an rapid, visuable method to detect the genotypes of HBV.Methods According to the published full-length sequence with known genotypes in GenBank, the specific primer and biotin-lablled probe were designed. We established a nucleic acid strip method for genotyping hepatitis B virus(HBV) isolates among 150 HBV infected patients and 20 healthy controls, and compared the results with those obtained by real-time PCR. Results There was 34.00% genotype B , 61.33% genotype C, 4.00% genotype B/C in 150 samples, respectively, while the remaining 0.67% unknown. The results for this assay were high comparable to the method of real-time PCR. Conclusion With the similar sensitivity and specifity when compared with real-time PCR, this rapid method is suitable to the clinical setting.
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Objective To develop a rapid, accurate, specific method to detect causative agent of hand, foot and mouth disease (HFMD). Methods Specific primers and probe were designed based on highly conserved VP1 region of enterovirus 71, coxsackie virus A16 and enterovirus. The sensitivity and specificity of the real-time RT-PCR was evaluated with 35 stool samples collected from pediatric patients with suspected HFMD and 20 clinical samples from health pediatric patients. Results Out of 35 clinical samples from suspected HFMD, 35 samples were identified as positive for enterovirus, 25 clinical samples were identified as positive for enterovirus 71, 8 clinical samples were identified as positive for coxsackie virus A16, among which 3 clinical samples were identified as positive for enterovirus 71 and coxsackie virus A16. The clinical diagnostic accordance rate is 85.71%. Out of 20 clinical samples from normal pediatric patients, 5 clinical samples were identified as positive for enterovirus, 20 clinical samples were negative for enterovirns 71 and coxsackie virus AI6. Conclusion Our results indicate real-time RT-PCR offers a rapid, sensitive, specific and cheap method to detect pathogen of HFMD from clinical specimens.