Performance evaluation of the VERSANT HCV RNA qualitative assay by using transcription-mediated amplification
Performance evaluation of the VERSANT HCV RNA qualitative assay by using transcription-mediated amplification
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DOI：
10.1128/jcm.41.1.310-317.2003
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发表时间：
2003-01-01
影响因子：
9.4
通讯作者：
Comanor, L
中科院分区：
文献类型：
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作者：
Gorrin, G;Friesenhahn, M;Comanor, L
根据美国国家临床实验室标准委员会和美国食品和药物管理局的指导方针，对转录介导扩增(TMA)检测丙型肝炎病毒(HCV)RNA的定性检测方法进行了临床前评估。我们的结果表明，该方法对临床样本的实验表明，丙型肝炎病毒TMA以类似的效率检测所有丙型肝炎病毒基因型，在50份丙型肝炎病毒RNA拷贝/毫升时，检测到来自感染丙型肝炎病毒1a、2b、3a、4a、5a和6a的患者的样本中大于或等于95%的样本。在与RNA转录本的实验中，在50个丙型肝炎病毒RNA拷贝/毫升时，丙型肝炎病毒TMA检测到大于或等于96.6%的来自丙型肝炎病毒1a、1b、2a、2c、3a、4a、5a和6a的转录本。来自丙型肝炎病毒2b基因型的转录本的检测在50拷贝/毫升时略低(88.4%)，但在75拷贝/毫升时为97.0%。此外，在临床实验室常见的各种条件下，包括长期储存、多次冻融循环、不同的采集管以及存在内源性物质、常用处方药或其他微生物和病毒的情况下，丙型肝炎病毒TMA在检测样本中的丙型肝炎病毒RNA方面表现出强劲的性能。丙型肝炎病毒TMA具有较高的敏感性、特异性、重复性和等同的基因型反应性，可作为临床实验室常规丙型肝炎病毒RNA定性检测的替代方法。
A preclinical evaluation of a qualitative assay for the detection of hepatitis C virus (HCV) RNA by transcription-mediated amplification (TMA) was conducted according to the guidelines of the National Committee for Clinical Laboratory Standards and the U.S. Food and Drug Administration. Our results showed that this assay, HCV TMA, detected 95% of samples with HCV RNA concentrations of 5.3 IU/ml and 29 copies/ml. HCV TMA showed an overall specificity of 99.6% and was highly reproducible, detecting 99.3% of samples with HCV RNA concentrations of 50 copies/ml across seven different lots of reagents. Experiments with clinical samples showed that HCV TMA detected all HCV genotypes with similar efficiencies, detecting greater than or equal to95% of samples at 50 HCV RNA copies/ml from patients infected with HCV genotypes 1a, 2b, 3a, 4a, 5a, and 6a. In experiments with RNA transcripts, HCV TMA detected greater than or equal to96.6% of transcripts derived from HCV genotypes 1a, 1b, 2a, 2c, 3a, 4a, 5a, and 6a at 50 HCV RNA copies/ml. Detection of transcripts derived from HCV genotype 2b was slightly lower (88.4%) at 50 copies/ml but was 97.0% at 75 copies/mi. In addition, HCV TMA exhibited robust performance in detecting HCV RNA in samples subjected to various conditions commonly encountered in a clinical laboratory, including long-term storage, multiple freeze-thaw cycles, different collection tubes, and the presence of endogenous substances, commonly prescribed drugs, or other microorganisms and viruses. With its high sensitivity, specificity, reproducibility, and equivalent genotype reactivity, HCV TMA may provide an attractive alternative for routine qualitative HCV RNA testing in clinical laboratories.