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通过聚合酶链反应和限制性酶切分析快速鉴别汉滩病毒和汉城病毒

Rapid differentiation between Hantaan and Seoul viruses by polymerase chain reaction and restriction enzyme analysis.

作者信息

Kim E C, Kim I S, Choi Y, Kim S G, Lee J S

机构信息

Department of Clinical Pathology, Seoul National University College of Medicine, Korea.

出版信息

J Med Virol. 1994 Jul;43(3):245-8. doi: 10.1002/jmv.1890430309.

Abstract

The majority of causative strains of hemorrhagic fever with renal syndrome (HFRS) are known as Hantaan and Seoul viruses in Korea. The clinical manifestations may be indistinguishable between both viruses, although the clinical course of Hantaan virus infection is more severe than that of the Seoul virus. Therefore, the differentiation of Hantaan or Seoul virus may be important for predicting the prognosis. The primers were selected from the published sequences of the S segments of Hantaan virus strain 76-118 and Seoul virus strain SR-11, which made it possible to obtain the same size of 403 bp amplified product by reverse transcriptase polymerase chain reaction (RT-PCR) and nested PCR from both viral strains. The differentiation of the amplified products was carried out by restriction enzyme digestion. With HindIII, the 403 bp amplified product from Hantaan virus strain 76-118 was cleaved into two segments of 175 bp and 228 bp. By contrast, the 403 bp product from Seoul virus strain SR-11 was not cut by HindIII. With HinfI, the 403 bp amplified product from Hantaan virus strain 76-118 was divided into two bands of 280 bp and 60 bp on the electrophoresis. In the case of the digestion of 403 bp PCR product from Seoul virus strain SR-11 with HinfI, more than four bands (155 bp, 115 bp, 60 bp, and 32/29 bp) were observed on the 2% agarose gel electrophoresis. This rapid technique may be useful for the differential diagnosis of Asian HFRS in Korea.

摘要

在韩国,肾综合征出血热(HFRS)的大多数致病菌株被称为汉坦病毒和汉城病毒。尽管汉坦病毒感染的临床病程比汉城病毒更严重,但两种病毒的临床表现可能难以区分。因此,区分汉坦病毒和汉城病毒对于预测预后可能很重要。引物是从汉坦病毒株76 - 118和汉城病毒株SR - 11的S片段已发表序列中选择的,这使得通过逆转录聚合酶链反应(RT-PCR)和巢式PCR从两种病毒株中获得相同大小的403 bp扩增产物成为可能。扩增产物的区分通过限制性酶切进行。用HindIII酶切时,汉坦病毒株76 - 118的403 bp扩增产物被切割成175 bp和228 bp的两个片段。相比之下,汉城病毒株SR - 11的403 bp产物未被HindIII切割。用HinfI酶切时,汉坦病毒株76 - 118的403 bp扩增产物在电泳中被分为280 bp和60 bp的两条带。在用HinfI酶切汉城病毒株SR - 11的403 bp PCR产物时,在2%琼脂糖凝胶电泳上观察到超过四条带(155 bp、115 bp、60 bp和32/29 bp)。这种快速技术可能有助于韩国亚洲HFRS的鉴别诊断。

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