Detection and quantification of hepatitis B virus DNA by three different methods in HBsAg positive patients

The detection and quantification of hepatitis B virus genomes appear to be the most reliable methods for monitoring HBV infection and assessing response to antiviral treatment. The aim of this study assess the performance of three HBV DNA detection and quantification assays currently used for the management of HBV infected patients. In our study. 44 HBsAg +ve samples were assayed to HBV DNA detection and quantification by three different methods [In House HBV PCR. Amplicor HBV Monitor and HBV branched DNA DNA "bDNA"]. Twenty three samples [52.27%] were HBV DNA positive by one or more of the three methods and 21 samples [47.73 3/4] were HBV DNA negative by the three methods. Twenty one samples [47.73 3/4] were HBV DNA positive by Amplicor HBV Monitor and their cops number ranged from 440 - 40x106 copies/ml. Ten samples [22.73%] were HBV DNA positive by in House HBV PCR with copy number ranged from 6400 - 40 x 106 copies/ml. Eight samples [18.18%] were HBV DNA positive by NBV branched DNA [bDNA] and their copy number ranged from I.2x106 40x106 copies/ml. Branched DNA [bDNA] versus Amplicor HBV Monitor showed that: [i] bDNA and Arnplicor HBV Monitor gave concordant results for 27 [61.63%] of 44 the samples. The assays were both positive in six samples [13.64%]. The assays were both negative in 21 samples [47.73%]. [ii] bDNA and Amplicor HBV Monitor gave discordant results in 17 cases [38.64%]: bDNA was positive and Amplicor HBV Monitor negative in two cases [4.55%]. While bDNA was negative and Amplicor HBV Monitor positive in 15 cases [34.1%], HBV-DNA load in the Monitor assay was higher in 14] NA positive samples [mean = 9.7 x 106 copies/ml] than bDNA negative samples [mean 4.1 x 104 copies ml]. In House positive [mean = 9.7 x 106 copies/ml] than bDNA negative samples [mean 4.1 x 104 copies/ml]. In House PCR and Amplicor HBV Monitor showed that [i] In House PCR and Amplicor HBV Monitor gave concordant results with 33 of the 44 samples [75.0%]. The assays were both positive with to samples [22.73%]. The assays were both negative in 23 cases [52.27%] [ii] In House PCR and Amplicor HBV Monitor gave discordant results in 11 cases [25.0%] In House PCR was negative and Amplicor HBV Monitor gave discordant results in 11 casses [26.0%] in House PCR was negative and Amplicor HBV Monitor gave discordant resutlds in 11 cases [25.0%]. In House PCR was negative and Amplicor HBV Montor positive in these ii cases. HBV-DNA load in the Monitor assay was higher in In House PCR positive samples [mean 5.88 x 106 copies/ml] than In House PCR negative samples [mean 21 x 102 copies mb. In House PCR versus bDNA showed that: [i] in House PCR and bDNA gave concordant results with 38 of the 44 samples [86.36%]. The assays were both positive in six cases [13.64%]. The assays were both negative in 32 caes [72.73%]. [ii] In House PCR and bDNA gave discordant results in six cases [13.64%] In House PCR was positive and bDNA negative with four samples [0.09%]. In House PCR was negative and bDNA positive with two samples [4.55%]. HBV-DNA load in the Monitor assay was higher in bDNA positive samples [mean x 10 copies mb than In House PCR positive samples [mean 5.88 x 102 copies mb. The monitor assasy was more sensitive than both the In House PCR and bDNA. This better sensitivity appeared to be clinically relevant [even their respective performance, these three assays should be used in complementary fashion in the management of HBV infected patients

Evaluation of inhouse HBV DMA PCR, and its comparison with amplicor HBV PCR [Roche] and quantiplex HBV bDNA [Chiron] in detection of hepatitis b virus infection

To evaluate the diagnostic efficacy of inhouse HBV DNA PCR detected either by ELISA or agarose gel electrophoresis techniques this study was designed. 60 patients having either HbsAg antigen or HBc IgG or both were selected from the outpatients medical clinic and inpatient medical section of the National liver institute, Minoufiya University. Inhouse HBV DNA PCR detected either by ELISA or agarose gel electrophoresis techniques were done for those patients, and the results were expressed in comparison with the results of Amplicor HBV PCR [Roche], and quantiplex HBV bDNA [Chiron]. Out of sixty patients who were serologically positive for either HbsAg, anti- HBc IgG or both, the inhouse HBV DNA PCR detected by ELISA was positive in 11 patients [18.33%], while inhouse HBV DNA PCR detected by EB and gel electrophoresis was positive in 9 patients [15%], also, quantiplex HBV bDNA [Chiron] was positive in 8 [17.78%] out of 45 patients only and amplicor HBV DNA PCR [Roche] was positive in 28 [46.67%] out of 60 patients. Also, the study revealed that the sensitivity of inhouse HBV DNA PCR detected by ELISA was 39.28%, with 100% specificity and 71.67% diagnostic accuracy, while inhouse HBV DNA PCR detected by agarose gel electrophoresis after ethidium bromide staining showed 32.14% sensitivity, 100% specificity and 68.33% diagnostic accuracy also, Also, quantiplex HBV bDNA [Chiron] showed 28.57% sensitivity, 91.66% specificity and 62.22% diagnostic accuracy when compared with the positive results obtained with amplicor HBV PCR [Roche]. So, due to its low sensitivity in comparison with amplicor HBV DNA PCR [Roche] we can not rely on inhouse PCR detected either by ELISA or agarose gel electrophoresis after ethidium bromide staining in diagnosis of HBV infection, further optimization of inhouse PCR is recommended for good yield and better diagnosis