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一种用于检测腐败支原体的特异性聚合酶链反应,腐败支原体是山羊传染性无乳综合征的病原体之一。

A specific PCR for the detection of Mycoplasma putrefaciens, one of the agents of the contagious agalactia syndrome of goats.

作者信息

Peyraud A, Woubit S, Poveda J B, De la Fe C, Mercier P, Thiaucourt F

机构信息

CIRAD EMVT Sante Animale, Animal Health Program, TA30/G Campus International de Baillarguet, 34398 Montpellier, France.

出版信息

Mol Cell Probes. 2003 Dec;17(6):289-94. doi: 10.1016/j.mcp.2003.07.006.

Abstract

Mycoplasma putrefaciens is listed as one of the etiologic agents of the contagious agalactia syndrome by the world organisation for animal health. This species has been characterized only recently, 1974, and the number of outbreaks caused by this microorganism so far is very scarce. It induces mastitis in infected goats although other symptoms such as arthritis in adults and septicaemia in kids are also frequently described. Up to now, the identification of M. putrefaciens relied on classical isolation and identification techniques which present a number of limitations. Specific primers for PCR have been designed based on sequence comparisons of the ArcB gene among the 'Mycoplasma mycoides cluster' and related species such as Mycoplasma cottewii and Mycoplasma yeatsii. Sequence alignments confirmed the taxonomic position of M. putrefaciens, which is related to the 'M. mycoides cluster' but also very close to M. yeatsii. The polymorphism observed amongst the different ArcB sequences allowed the determination of a primer pair yielding a specific amplification of a 316 bp-long DNA fragment by PCR. This PCR was validated in two different laboratories with a variety of mycoplasma strains isolated from goats. This new PCR technique will be very useful for a quicker determination of M. putrefaciens strains as well as a better understanding of the prevalence of M. putrefaciens infections.

摘要

腐败支原体被世界动物卫生组织列为传染性无乳综合征的病原之一。该物种直到1974年才被鉴定出来,到目前为止由这种微生物引起的疫情非常罕见。它可导致感染山羊患乳腺炎,不过也经常出现其他症状,如成年羊患关节炎,幼羊患败血症。到目前为止,腐败支原体的鉴定依赖于传统的分离和鉴定技术,这些技术存在一些局限性。基于“丝状支原体簇”以及相关物种(如考氏支原体和耶氏支原体)中ArcB基因的序列比较,设计了用于PCR的特异性引物。序列比对证实了腐败支原体的分类地位,它与“丝状支原体簇”相关,但也与耶氏支原体非常接近。在不同的ArcB序列中观察到的多态性使得能够确定一对引物,通过PCR特异性扩增出一个316 bp长的DNA片段。该PCR在两个不同实验室用从山羊分离的多种支原体菌株进行了验证。这种新的PCR技术对于更快地鉴定腐败支原体菌株以及更好地了解腐败支原体感染的流行情况将非常有用。

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