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[通过连接酶B基因PCR和测序对致病性钩端螺旋体进行鉴别]

[Differentiation of pathogenic leptospires spp by PCR of ligB gene and sequencing].

作者信息

Martínez Mara Leila, Grune Loffler Sylvia, Romero Graciela Noemi, Brihuega Bibiana Felicitas

机构信息

Instituto de Patobiología, Centro de Investigación en Ciencias Veterinarias y Agronómicas (CICVyA), INTA Castelar, Hurlingham, Buenos Aires, Argentina.

Instituto de Patobiología, Centro de Investigación en Ciencias Veterinarias y Agronómicas (CICVyA), INTA Castelar, Hurlingham, Buenos Aires, Argentina.

出版信息

Rev Argent Microbiol. 2018 Apr-Jun;50(2):126-130. doi: 10.1016/j.ram.2016.11.008. Epub 2017 Oct 21.

Abstract

Leptospirosis is a zoonosis having worldwide distribution. The objective of this work was to develop a molecular technique to differentiate pathogenic Leptospira spp. A region of adhesin ligB, present only in the pathogenic species was amplified by PCR and sequenced. ligBRpet and ligBFpet primers were used, which amplified the target DNA from pathogenic L. interrogans reference strains serovars Pomona strain Pomona, Canicola strain Hond Utrecht IV, Copenhageni strain M 20, Wolffi strain 3705, Pyrogenes strain Salinem, Hardjo strain Hardjoprajitmo, L. borgpetersenii serovar Castellonis strain Castellon 3 and 4 pathogenic strains isolated from bovines, pigs, rats and opossums. L. biflexa serovars Patoc strain Patoc I and Andamana strain Andamana were not amplified. Sequencing of the amplified products exhibited sufficient variation among serovars, which differentiates them.

摘要

钩端螺旋体病是一种分布于全球的人畜共患病。这项工作的目的是开发一种分子技术来区分致病性钩端螺旋体属物种。通过PCR扩增并测序了仅存在于致病物种中的粘附素ligB区域。使用了ligBRpet和ligBFpet引物,它们从致病性问号钩端螺旋体参考菌株波摩那血清型波摩那菌株、犬型血清型洪德乌得勒支IV菌株、哥本哈根血清型M 20菌株、沃尔夫血清型3705菌株、致热血清型萨利内姆菌株、哈焦血清型哈焦普拉吉特莫菌株、博氏钩端螺旋体血清型卡斯泰洛尼血清型卡斯泰洛尼3菌株以及从牛、猪、大鼠和负鼠中分离出的4株致病菌株中扩增出目标DNA。双曲钩端螺旋体血清型帕托克血清型帕托克I菌株和安达马纳血清型安达马纳菌株未被扩增。扩增产物的测序显示血清型之间存在足够的差异,从而能够区分它们。

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