TYPING OF HANTAVIRUSES FROM 5 CONTINENTS BY POLYMERASE CHAIN-REACTION
TYPING OF HANTAVIRUSES FROM 5 CONTINENTS BY POLYMERASE CHAIN-REACTION
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DOI：
10.1016/0168-1702(92)90142-v
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发表时间：
1992-10-01
期刊：
影响因子：
5
通讯作者：
KANG, CY
中科院分区：
文献类型：
--
作者：
PUTHAVATHANA, P;LEE, HW;KANG, CY
汉坦病毒是布尼亚病毒科的一个属，包括至少四种血清学上不同的类型：汉坦病毒、首尔病毒、普马拉病毒和前景山病毒。本文报告了用聚合酶链反应（PCR）对来自5个不同大陆的27个独立分离的汉坦病毒进行分型。用酸性硫氰酸胍-苯酚-氯仿法从病毒感染的Vero E6细胞单层中提取细胞总RNA。我们使用了5组不同的寡核苷酸引物，长度范围从18到22个核苷酸;一组特异于S基因组片段的保守区域并用作属特异性引物，另外4组引物是从M基因组片段的独特序列设计的，使得每个引物组仅特异于一种血清学类型的汉坦病毒。PCR产物经限制性内切酶酶切鉴定。在汉滩、首尔、普马拉和前景山分别有10、12、3和1株分离株。PCR与血清学分型结果符合率为100%，可作为血清学方法的辅助试验或作为汉坦病毒新分离株的诊断和分型的独立试验。
Hantavirus, a genus in the family Bunyaviridae, is comprised of at least four serologically distinct types: Hantaan, Seoul, Puumala and Prospect Hill. The present communication reports the use of polymerase chain reaction (PCR) for typing 27 independently isolated Hantaviruses from 5 different continents. Total cellular RNA was extracted from virus-infected Vero E6 cell monolayers by the acid guanidium thiocyanate-phenol-chloroform method. We have utilized 5 different sets of oligonucleotide primers ranging from 18 to 22 nucleotides in length; one set was specific for a conserved region of the S genomic segment and used as genus-specific primers, the other 4 sets of primers were designed from unique sequences of the M genomic segment such that each primer set was specific to only one serological type of Hantavirus. The PCR products were analyzed by restriction endonuclease digestion for further confirmation. We typed 10, 12, 3 and 1 isolates into Hantaan, Seoul, Puumala and Prospect Hill respectively. The results of PCR were 100% agreeable with that of serological typing, and thus, PCR can be used as an adjunct test with serological method(s) or an independent test for diagnosis and for typing of new isolates of Hantaviruses.