Künkel U, Höhne M, Berg T, Hopf U, Kekulé A S, Frösner G, Pauli G, Schreier E
Robert Koch-Institute, Berlin, Germany.
J Hepatol. 1998 Jun;28(6):978-84. doi: 10.1016/s0168-8278(98)80346-2.
BACKGROUND/AIMS: A novel virus, GBV-C/HGV, with a genome RNA organization similar to that of the Flaviviridae family was identified in sera of patients with hepatitis. The presence of GBV-C/HGV RNA can only be determined by the amplification of genomic regions using the reverse transcriptase-polymerase chain reaction (RT-PCR).
To assess the quality of the RT-PCR, 14 laboratories investigated a coded serum panel that comprised three GBV-C/HGV RNA-positive sera from three different patients, dilutions of these sera, and three GBV-C/HGV RNA-negative serum samples, two of which were collected from patients with hepatitis C but without GBV-C/HGV infection. In-house RT-PCR as well as commercially available GBV-C/HGV test kits were used in this study.
Only four laboratories (29%) reported the expected results, and four laboratories (29%) false-positive results; nine laboratories (64%) reported at least one false-negative result. Eleven laboratories (79%) detected the undiluted samples. The majority of false results were obtained with the dilutions of GBV-C/HGV RNA-positive samples. Negative results in the 10(-4) dilution were not considered to be false-negative, since during pre-screening GBV-C/HGV RNA had been detected in this dilution in only 50% of assays by the three laboratories involved in organizing the evaluation. Results obtained by commercial kits and by in-house assays were indiscriminate in quality of performance in this study.
To facilitate further quality assessment studies on the performance of GBV-C/HGV RNA detection, an international GBV-C/HGV RNA standard should be made available. Further efforts are required to optimize GBV-C/HGV RT-PCR.
背景/目的：在肝炎患者血清中发现了一种新型病毒，GBV-C/HGV，其基因组RNA结构与黄病毒科相似。GBV-C/HGV RNA的存在只能通过逆转录聚合酶链反应（RT-PCR）扩增基因组区域来确定。
为评估RT-PCR的质量，14个实验室对一个编码血清样本进行了检测，该样本包括来自三名不同患者的三份GBV-C/HGV RNA阳性血清、这些血清的稀释液以及三份GBV-C/HGV RNA阴性血清样本，其中两份来自丙型肝炎患者但未感染GBV-C/HGV。本研究使用了内部RT-PCR以及市售的GBV-C/HGV检测试剂盒。
只有四个实验室（29%）报告了预期结果，四个实验室（29%）报告了假阳性结果；九个实验室（64%）报告了至少一个假阴性结果。11个实验室（79%）检测了未稀释的样本。大多数错误结果是在GBV-C/HGV RNA阳性样本的稀释液检测中获得的。10^(-4)稀释度的阴性结果不被视为假阴性，因为在预筛查中，参与组织评估的三个实验室仅在50%的检测中在该稀释度下检测到GBV-C/HGV RNA。在本研究中，市售试剂盒和内部检测所获得的结果在性能质量上没有区别。
为便于对GBV-C/HGV RNA检测性能进行进一步的质量评估研究，应提供国际GBV-C/HGV RNA标准。需要进一步努力优化GBV-C/HGV RT-PCR。
