Eiden J J, Wilde J, Firoozmand F, Yolken R
Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
J Clin Microbiol. 1991 Mar;29(3):539-43. doi: 10.1128/jcm.29.3.539-543.1991.
A combined reverse transcriptase reaction-polymerase chain reaction (RT-PCR) was developed to achieve the sensitive detection of group B rotaviruses (GBR). Sequences derived from genomic segment 3 of the IDIR (intestinal disease of infant rats) strain of GBR permitted the detection of greater than or equal to 0.08 pg of purified IDIR genomic RNA (4,000 genome copies). Primers complementary to the terminal sequences of gene 11 of GBR strain ADRV (adult diarrhea rotavirus) allowed for the detection of as little as 0.008 pg of purified ADRV genomic RNA. Detection of heterologous strains of GBR was also observed with these primer pairs. IDIR gene 3 primers recognized greater than or equal to 8 pg of RNA from bovine GBR obtained from a variety of geographic locations. RNA from IDIR, but not bovine GBR, strains was detected by means of RT-PCR with ADRV gene 11 primers. Neither set of GBR primers was reactive in RT-PCR with fecal specimens containing group A rotaviruses or fecal specimens from uninfected controls. This RT-PCR assay permits the sensitive and specific detection of a variety of GBR in fecal specimens.
为实现对B组轮状病毒(GBR)的灵敏检测,开发了一种逆转录反应-聚合酶链反应(RT-PCR)联合方法。源自GBR的IDIR(幼鼠肠道疾病)株基因组片段3的序列能够检测到≥0.08 pg的纯化IDIR基因组RNA(4000个基因组拷贝)。与GBR株ADRV(成人腹泻轮状病毒)基因11末端序列互补的引物能够检测低至0.008 pg的纯化ADRV基因组RNA。使用这些引物对也观察到了对GBR异源株的检测。IDIR基因3引物能够识别来自不同地理位置的牛GBR中≥8 pg的RNA。通过使用ADRV基因11引物的RT-PCR检测到了IDIR株而非牛GBR株的RNA。两组GBR引物在对含A组轮状病毒的粪便标本或未感染对照的粪便标本进行RT-PCR时均无反应。这种RT-PCR检测方法能够灵敏且特异检测粪便标本中的多种GBR。