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基于墨西哥狂犬病病毒变种分子分型开发引物对

Development of Primer Pairs from Molecular Typing of Rabies Virus Variants Present in Mexico.

作者信息

Bastida-González Fernando, Ramírez-Hernández Dolores G, Chavira-Suárez Erika, Lara-Padilla Eleazar, Zárate-Segura Paola

机构信息

Laboratorio de Medicina Traslacional, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, Santo Tomás, Miguel Hidalgo, 11340 Ciudad de México, CDMX, Mexico; Laboratorio de Biología Molecular, Laboratorio Estatal de Salud Pública del Estado de México, Paseo Tollocan s/n, La Moderna de la Cruz, 50180 Toluca, MEX, Mexico.

Laboratorio de Biología Molecular, Laboratorio Estatal de Salud Pública del Estado de México, Paseo Tollocan s/n, La Moderna de la Cruz, 50180 Toluca, MEX, Mexico.

出版信息

Biomed Res Int. 2016;2016:4659470. doi: 10.1155/2016/4659470. Epub 2016 Aug 3.

Abstract

Nucleoprotein (N) gene from rabies virus (RABV) is a useful sequence target for variant studies. Several specific RABV variants have been characterized in different mammalian hosts such as skunk, dog, and bats by using anti-nucleocapsid monoclonal antibodies (MAbs) via indirect fluorescent antibody (IFA) test, a technique not available in many laboratories in Mexico. In the present study, a total of 158 sequences of N gene from RABV were used to design eight pairs of primers (four external and four internal primers), for typing four different RABV variants (dog, skunk, vampire bat, and nonhematophagous bat) which are most common in Mexico. The results indicate that the primer and the typing variant from the brain samples, submitted to nested and/or real-time PCR, are in agreement in all four singleplex reactions, and the designed primer pairs are an alternative for use in specific variant RABV typing.

摘要

狂犬病病毒(RABV)的核蛋白(N)基因是变异研究中一个有用的序列靶点。通过间接荧光抗体(IFA)试验,利用抗核衣壳单克隆抗体(MAb),已在臭鼬、狗和蝙蝠等不同哺乳动物宿主中鉴定出几种特定的RABV变异株,而墨西哥的许多实验室都没有这种技术。在本研究中,共使用了158个RABV的N基因序列来设计八对引物(四对外引物和四对内引物),用于对墨西哥最常见的四种不同RABV变异株(狗、臭鼬、吸血蝙蝠和非吸血蝙蝠)进行分型。结果表明,在所有四个单重反应中,用于脑样本的引物和分型变异株,经巢式和/或实时PCR检测,结果一致,且所设计的引物对可作为特定变异RABV分型的替代方法。

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