通过修正实时 RT-PCR 检测中正向引物的三个错配核苷酸，提高了对各种轮状病毒基因型在粪便标本中的检测敏感性。
Increased sensitivity for various rotavirus genotypes in stool specimens by amending three mismatched nucleotides in the forward primer of a real-time RT-PCR assay.
机构信息
Provincial Public Health Laboratory, Edmonton, Alberta, Canada.
出版信息
J Virol Methods. 2011 Mar;172(1-2):85-7. doi: 10.1016/j.jviromet.2010.12.013. Epub 2010 Dec 23.
The real-time TaqMan RT-PCR assay (Pang et al., 2004) did not detect 14 clinical samples with rotavirus G2 genotype. Three to five nucleotides (nt) were found to be mismatched between the published forward primer when compared to G2P[4], G2P[8], G3P[4], G9P[4], G8 and G12 sequences. An additional forward primer was designed and included in a modified assay to test the 14 clinical samples and 12 samples with known rotavirus G and P genotypes. The modified assay has improved significantly the sensitivity for specific rotavirus strains without affecting the detection of other genotypes, creating a molecular assay with broad detection of various genotypes of group A rotaviruses.
实时 TaqMan RT-PCR 检测法（Pang 等人，2004 年）未能检测到 14 份轮状病毒 G2 基因型的临床样本。与 G2P[4]、G2P[8]、G3P[4]、G9P[4]、G8 和 G12 序列相比，发表的正向引物与发现的 3 到 5 个核苷酸（nt）不匹配。设计了一个额外的正向引物并包含在一个改良的检测方法中，以检测 14 份临床样本和 12 份已知的轮状病毒 G 和 P 基因型样本。改良的检测方法显著提高了特定轮状病毒株的检测灵敏度，而不会影响其他基因型的检测，创建了一种具有广泛检测 A 组轮状病毒各种基因型的分子检测方法。
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