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首页> 外文期刊>Clinical gastroenterology and hepatology: the official clinical practice journal of the American Gastroenterological Association >Commercial real-time reverse transcriptase PCR assays can underestimate or fail to quantify hepatitis delta virus viremia.
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Commercial real-time reverse transcriptase PCR assays can underestimate or fail to quantify hepatitis delta virus viremia.

机译：商业实时逆转录酶PCR检测方法可能会低估或无法量化肝炎三角洲病毒的病毒血症。

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摘要

Hepatitis delta virus (HDV) infection causes fulminant hepatitis and increases the severity of chronic hepatitis B virus infection, leading to cirrhosis, liver failure, or hepatocellular carcinoma. There are 8 HDV genotypes (genotypes 1-8). We previously developed a TaqMan real-time reverse transcriptase (RT)-PCR method that is able to quantify viral load of all HDV genotypes (linear from 2 to 8 log(10) copies/mL). We compared its results with those from 3 commercial real-time RT-PCR assays: the Lightmix HDV kit (designed to quantify HDV genotype 1 [HDV-1]), and the RoboGene and the DiaPro HDV RNA quantification kits (designed to quantify all genotypes).We selected RNA from 128 clinical samples of all HDV genotypes except HDV-4, with various HDV viral load values. We also analyzed 5 samples, collected over time, from each of 6 patients infected with strains of different genotypes.Quantification results from the commercial kits for HDV-1 from European or Asian samples were consistent with those from our method, however, they underestimated (0.5-1 log(10) with Lightmix and DiaPro) and did not detect (1 and 4 samples with Lightmix and DiaPro, respectively) HDV-1 African samples. Moreover, the commercial kits greatly underestimated HDV viral load of almost all non-genotype-1 strains (about 2-3 log(10)), and even did not detect HDV-7 or HDV-8 RNA in several samples with high concentrations of virus.Commercial kits accurately quantify HDV-1 in samples from European and Asian patients. However, they can dramatically underestimate or fail to quantify HDV viral load from samples from African patients infected with strains of genotypes 1 and 5 to 8.
机译：三角洲肝炎病毒（HDV）感染会导致暴发性肝炎，并增加慢性乙型肝炎病毒感染的严重性，从而导致肝硬化，肝衰竭或肝细胞癌。 HDV基因型有8种（基因型1-8）。我们之前开发了TaqMan实时逆转录酶（RT）-PCR方法，该方法能够量化所有HDV基因型的病毒载量（线性范围为2至8 log（10）拷贝/ mL）。我们将其结果与3种商业实时RT-PCR分析的结果进行了比较：Lightmix HDV试剂盒（旨在量化HDV基因型1 [HDV-1]），以及RoboGene和DiaPro HDV RNA定量试剂盒（旨在量化所有我们从128种除HDV-4以外的所有HDV基因型的临床样本中选择了RNA，具有各种HDV病毒载量值。我们还分析了6例感染不同基因型菌株的患者中随时间推移收集的5个样本。欧洲或亚洲样本的商业化HDV-1试剂盒的定量结果与我们的方法一致，但是，他们低估了（使用Lightmix和DiaPro的检测结果为0.5-1 log（10）），未检测到（分别使用Lightmix和DiaPro的检测结果为1和4个样品）HDV-1非洲样品。而且，这种商用试剂盒大大低估了几乎所有非基因型1菌株的HDV病毒载量（约2-3 log（10）），甚至没有检测到高浓度的HDV-7或HDV-8 RNA样本。商业试剂盒可准确定量欧洲和亚洲患者样本中的HDV-1。但是，它们可能会大大低估或无法量化来自感染了基因型1和5至8的非洲患者的样本中的HDV病毒载量。

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