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目的 通过病毒粒 7个不同RNA节段核苷酸序列测定和分析 ,进一步排除A/广东 / 6 /91(H1N1)毒株由实验室污染而来。它与A/PR/ 8/ 34 (H1N1)毒株基因组间有哪些RNA节段存在有差异。同时它是否是一株基因重配株 ?方法 病毒粒RNA经逆转录合成cDNA ,经聚合酶链反应 (PCR)扩增 ,产物纯化 ,采用双脱氧链末端终止法进行核苷酸序列测定。结果 所比较的 7个不同RNA节段中 ,测定毒株RNA6和 7的核苷酸序列与A/PR/ 8/ 34 (H1N1)完全相同。而RNA1,2 ,3,5和 8间核苷酸差异数分别为 2 0 ,5 ,11,7和 6。这些差异导致了所编码的PB2 ,PB1,PA ,NP和NS蛋白分子上氨基酸序列差异数分别为 10 ,2 ,1,0和 4。结论 A/广东 / 6 / 91(H1N1)毒株由A/PR/ 8/ 34 (H1N1)病毒实验室污染而来可完全加以排除。它是A/PR/ 8/ 34 (H1N1)病毒类似毒株 ,而不是基因重配株。
Objective To further exclude the contamination of laboratory A / Guangdong / 6/91 (H1N1) strains by the determination and analysis of nucleotide sequences of seven different RNA segments of virion. There are differences in RNA segments between it and the genome of strain A / PR / 8/34 (H1N1). At the same time, is it a gene reassortant? Methods The virus RNA was reverse transcribed into cDNA, amplified by polymerase chain reaction (PCR) and the product was purified. The nucleotide sequence was determined by dideoxy chain termination method. Results Among the 7 different RNA segments compared, the nucleotide sequences of the strains RNA6 and 7 were determined to be identical to A / PR / 8/34 (H1N1). While the number of nucleotide differences between RNAs 1, 2, 3, 5, and 8 is 20, 5, 11, 7, and 6, respectively. These differences result in amino acid sequence differences of 10, 2, 1, 0 and 4 for the encoded PB2, PB1, PA, NP and NS protein molecules, respectively. Conclusion The A / Guangdong / 6/91 (H1N1) strain was completely eliminated from the A / PR / 8/34 (H1N1) virus laboratory. It is a similar strain of A / PR / 8/34 (H1N1) virus, not a genetic reassortant.