Detection and differentiation of human hepatitis A strains by commercial quantitative real-time RT-PCR tests.
Performance characteristics of two commercial quantitative Hepatitis A virus (HAV) RT-PCR assays, the LightCycler Hepatitis A virus quantification kit (Roche Diagnostics) and the RealArt HAV LC RT PCR kit (artus GmbH) for detection and quantification of HAV were evaluated. Both assays rely on reverse transcription and amplification of extracted RNA. Dilutions of two HAV strains, belonging to different subtypes, were prepared to determine the precision, accuracy, linearity and the detection limit. Both assays were found to be suitable for quantification measurement of HAV RNA, but only the Roche kit was able to distinguish the different HAV strains tested. The linear range for the artus assay was 10(4)-10(6)IU/ml and 2 x 10(4) to 2 x 10(8) RNA copies/ml for the Roche assay. The detection limit of Roche kit was 2 TCID(50)/ml or 500 RNA copies/ml and 5 TCID(50)/ml or 50 IU/ml for the artus kit. Despite these small differences it is concluded that both assays are very suitable for detection and quantification of most prevalent HAV subtypes.
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Quality and Safety Assurance Department, Nestlé Research Center, Vers-chez-les-Blanc, CH-1000 Lausanne 26, Switzerland. gloria.sanchez-moragas@rdls.nestle.com
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D030041:Hepatitis A virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D011933:Reagent Kits, Diagnostic; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic