Development and evaluation of an improved diagnostic PCR for Mycoplasma synoviae using primers located in the haemagglutinin encoding gene vlhA and its value for strain typing
Development and evaluation of an improved diagnostic PCR for Mycoplasma synoviae using primers located in the haemagglutinin encoding gene vlhA and its value for strain typing
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DOI：
10.1016/j.vetmic.2008.10.011
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发表时间：
2009-04-14
影响因子：
3.3
通讯作者：
Bradbury, J. M.
中科院分区：
文献类型：
--
作者：
Hammond, P. P.;Ramirez, A. S.;Bradbury, J. M.
利用已发表的引物，对滑膜支原体进行检测，并利用vlhA基因序列进行菌株鉴定。然而，在21株滑膜分枝杆菌中，有3株无法扩增，因此设计了一种新的反向引物，目标位于vlhA基因的保守区域。这使得所有21株滑膜分枝杆菌菌株、另外9株菌株以及来自滑膜分枝杆菌阳性鸟类的11个拭子样本的材料都能产生PCR产物，表明该方法也适用于临床标本。然后在滑膜支原体型菌株和其他22种已知的禽支原体物种上对该方案进行了测试，仅扩增了滑膜支原体。进一步的测试表明，该PCR与用于检测滑膜分枝杆菌的其他PCR测试相同或更敏感。随后基于相似性百分比和进化关系的PCR产物DNA序列分析似乎是菌株分化的有用工具。(C) 2008 Elsevier B.V.版权所有
Using published primers, detection of Mycoplasma synoviae and strain identification using the vlhA gene sequence was attempted. However, of 21 M. synoviae strains examined three, could not be amplified, so a new reverse primer was designed with a target in the conserved region of the vlhA gene. This allowed all 21 M. synoviae strains, a further nine strains and also material from 11 swab samples from M. synoviae-positive birds, to produce a PCR product, suggesting that the method could also be suitable for clinical specimens. The protocol was then tested on the type strains of M. synoviae and the other 22 recognised avian Mycoplasma species, with amplification of M. synoviae only. Further testing demonstrated that this PCR was equally or more sensitive than other PCR tests used to detect M. synoviae. Subsequent DNA sequence analysis of the PCR product based on percent similarity and evolutionary relationship appeared to be a useful tool for strain differentiation. (C) 2008 Elsevier B.V. All rights reserved.