Detection and Genotyping of Hantaviruses Using Reverse Transcription·Polym erase Chain Reaction

LEI Wan-Li; TANG Jia-Qi; LU Cheng-Beng; PAN Xiu-Zhen; LI Xian-Fu

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February 1998

LEI Wan-Li, TANG Jia-Qi, LU Cheng-Beng, PAN Xiu-Zhen and LI Xian-Fu. Detection and Genotyping of Hantaviruses Using Reverse Transcription·Polym erase Chain Reaction[J]. Virologica Sinica, 1998, 13(1).

Citation: LEI Wan-Li, TANG Jia-Qi, LU Cheng-Beng, PAN Xiu-Zhen, LI Xian-Fu. Detection and Genotyping of Hantaviruses Using Reverse Transcription·Polym erase Chain Reaction .VIROLOGICA SINICA, 1998, 13(1) : 28.

肾综合征出血热病毒基因检测及分型的研究

1.
南京军区军事医学研究所南京农业大学动物医学院

摘要

根据流行于我国的两型ＨＦＲＳＶ代表株汉滩型７６１１８株及汉城型Ｒ２２株Ｍ节段的核酸序列，设计两型共同引物，建立了逆转录－聚合酶链反应（ＲＴＰＣＲ）方法，检测３９株从不同地区、不同宿主分离的ＨＦＲＳＶ感染鼠脑及细胞培养物；同时还建立了捕捉ＥＬＩＳＡ法（ｃＥＬＩＳＡ），检测了３９株中的３６株，每份样本设复孔，以Ｐ／Ｎ≥２．１０且Ｐ－Ｎ≥０．１０者判为阳性。ＲＴＰＣＲ及ｃＥＬＩＳＡ两法的检出率分别为９７．６％与８２．４％，二者符合率８４．６％。此外，对ＲＴＰＣＲ产物进行酶切分型，３８份扩增产物中的１５份可被ＡｌｕＩ切开。根据所获酶切图谱的差异，可分为汉滩型及汉城型两型，显示了酶切分型的潜在价值
- 肾综合征出血热病毒逆转录-聚合酶链反应捕捉ELISA法检测分型

Detection and Genotyping of Hantaviruses Using Reverse Transcription·Polym erase Chain Reaction

1.
East-China Medical Istitute of Biological Techniques，Nanjing 210002College of Veterinary Medicine，Nanjing Agrieultural University．Najing 210095

Abstract

Common primers were designed, based on M segmental sequences of two HFRSV serotypes—HTNV and SEOV, and RT PCR (reverse transcription-polymerase chain reaction) method was applied in detecting 39 strains of isolated hosts from different areas. Thirty six of the 39 samples were tested by cELISA (capture ELISA) in duplicated wells, while P/N≥2.10 and P-N≥0.10 the result was considered to the positive. The detection rates of RT PCR and cELISA were 97.6% and 82.4% respectively, 84.6% in correspondence. Fifteen of 38 RT PCR products could be digested by restriction endonuclease AluI. According to the restriction map the HFRSV detected was classified into two types——HTNV and SEOV.
- HFRSV RT PCR cELISA Detection Type

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Detection and Genotyping of Hantaviruses Using Reverse Transcription·Polym erase Chain Reaction

1. East-China Medical Istitute of Biological Techniques，Nanjing 210002College of Veterinary Medicine，Nanjing Agrieultural University．Najing 210095

Abstract: Common primers were designed, based on M segmental sequences of two HFRSV serotypes—HTNV and SEOV, and RT PCR (reverse transcription-polymerase chain reaction) method was applied in detecting 39 strains of isolated hosts from different areas. Thirty six of the 39 samples were tested by cELISA (capture ELISA) in duplicated wells, while P/N≥2.10 and P-N≥0.10 the result was considered to the positive. The detection rates of RT PCR and cELISA were 97.6% and 82.4% respectively, 84.6% in correspondence. Fifteen of 38 RT PCR products could be digested by restriction endonuclease AluI. According to the restriction map the HFRSV detected was classified into two types——HTNV and SEOV.