[Evaluation of the agreement of results obtained from ABI Prism 7000 and 7700 sequencers in the quantification of hepatitis B virus DNA].
There are different methods and systems for quantification of HBV-DNA in clinical virology laboratories. The aim of this study was to evaluate the agreement of the polymerase chain reaction (PCR) protocol with ABI Prism 7000 instrument (PE Biosystems) which was designed and optimised for ABI Prism 7700 (PE Biosystems). Serum samples obtained from 168 chronic hepatitis B patients were treated with "High Pure Viral Nucleic Acid Kit" (Roche Applied Science, USA), and MagnaPure LC isolation station (Roche Applied Science, Germany) was used for HBV-DNA isolation. Real time PCR procedure which amplifies pre-S gene of HBV genome was performed. Amplification and detection steps of all samples were performed with ABI Prism 7700 and 7000 Sequence Detection Systems. Among 168 samples, results of 124 serum samples were found to be in dynamic ranges of the tests. The results of these 124 samples obtained from ABI 7000 and ABI 7700 were concordant. Among the rest of 44 samples; one yielded higher than 10(10) copies/mL with two of the systems; six samples gave results higher than 10(10) copies/mL only with 7700; thirty samples were found negative with both of the systems; seven samples were positive (320-1220 copies/mL) with 7000 but negative with 7700. As a result this PCR protocol can be used in ABI 7000 system according to viral quality control (VQC) results. However, since the results of samples with HBV-DNA less than 1 x 10(6) copy/ml were discordant with the results obtained by ABI 7700 system, it can be concluded that different systems must not be used for the management and monitoring of the same patient.
Sertöz
R Yazan
RY
Ege Universitesi Tip Fakültesi, Mikrobiyoloji ve Klinik Mikrobiyoloji Anabilim Dali, Izmir.
Erensoy
S
S
Pas
S
S
Niesters
H G M
HG
D004279:DNA, Viral / Q000097:blood* / Q000302:isolation & purification; D006515:Hepatitis B virus / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000097:blood / Q000175:diagnosis / Q000821:virology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D011786:Quality Control; D015203:Reproducibility of Results
D004279:DNA, Viral