Muerhoff A S, Simons J N, Erker J C, Desai S M, Mushahwar I K
Virus Discovery Group, Experimental Biology Research, Abbott Laboratories, North Chicago, IL USA.
J Virol Methods. 1996 Oct;62(1):55-62. doi: 10.1016/0166-0934(96)02088-5.
Recently, the discovery of a new human RNA virus, GB virus C (GBV-C), was reported. GBV-C was isolated from the serum of a West African individual using degenerate oligonucleotide PCR primers designed from a consensus sequence of the NS3 helicase genes of hepatitis C virus (HCV), GBV-A, and GBV-B. Seven other individuals were shown to be infected with GBV-C via RT-PCR using these primers. Subsequently, degenerate PCR primers based upon a consensus sequence of the eight original isolates were designed. These primers were shown to be superior to the original set. However, since they were derived from a region of the viral genome exhibiting up to 17% nucleotide sequence divergence, mismatch between the primers and template may result in an underestimation of the true GBV-C prevalence. To overcome this potential problem, we obtained the sequences at the 5'-untranslated region (UTR) of the GBV-C genome from 35 infected individuals and identified regions of high sequence conservation among the isolates. We describe the design and testing of PCR primers derived from conserved sequences within the 5'-UTR of the GBV-C genome. These primers were shown to be as effective as the helicase-derived primers in detecting GBV-C RNA in human sera.
最近，有报道称发现了一种新的人类RNA病毒，即GB病毒C（GBV-C）。利用根据丙型肝炎病毒（HCV）、GBV-A和GBV-B的NS3解旋酶基因的共有序列设计的简并寡核苷酸PCR引物，从一名西非个体的血清中分离出了GBV-C。使用这些引物通过RT-PCR检测发现，另外7名个体也感染了GBV-C。随后，根据最初8个分离株的共有序列设计了简并PCR引物。结果显示这些引物比最初的引物更具优势。然而，由于它们源自病毒基因组中核苷酸序列差异高达17%的区域，引物与模板之间的错配可能会导致对GBV-C真实流行率的低估。为了克服这一潜在问题，我们从35名感染个体中获取了GBV-C基因组5'-非翻译区（UTR）的序列，并确定了分离株之间高度保守的序列区域。我们描述了源自GBV-C基因组5'-UTR保守序列的PCR引物的设计和测试。结果显示，这些引物在检测人血清中的GBV-C RNA方面与解旋酶衍生引物同样有效。
