|本期目录/Table of Contents|

[1]徐彬,张敬峰,卢凤英,等.基于特有基因VY93_RS02885为靶标的滑液支原体qPCR检测方法建立[J].江苏农业科学,2021,49(19):183-189.
　Xu Bin,et al.Establishment of a qPCR detection method for Mycoplasma synoviae based on unique gene VY93_RS02885 as the target[J].Jiangsu Agricultural Sciences,2021,49(19):183-189.
点击复制

基于特有基因VY93_RS02885为靶标的滑液支原体qPCR检测方法建立(PDF)
分享到：

《江苏农业科学》[ISSN:1002-1302/CN:32-1214/S]

卷:
第49卷
期数:
2021年第19期
页码:
183-189
栏目:
畜牧兽医与水产蚕桑
出版日期:
2021-10-05

文章信息/Info

Title:
Establishment of a qPCR detection method for Mycoplasma synoviae based on unique gene VY93_RS02885 as the target
作者:
徐彬; 张敬峰; 卢凤英; 孙华伟; 赵莎; 张晓曦; 刘青涛; 张小飞
江苏省农业科学院兽医研究所/农业部兽医生物工程技术重点实验室，江苏南京 210014
Author(s):
Xu Bin; et al
关键词:
滑液支原体; 特有基因; qPCR; 检测方法
Keywords:
-
分类号:
S852.62
DOI:
-
文献标志码:
A
摘要:
利用CD-HIT-EST快速聚类分析和BLASTN同源性比对共鉴定出17个滑液支原体(Mycoplasma synoviae)特有基因，并从中选取保守基因VY93_RS02885作为分子诊断的靶标基因。根据该基因核苷酸序列设计了1对引物，建立了滑液支原体的SYBR green qPCR检测方法。结果表明，该方法对不同浓度的模板和对应的CT值具有较好的线性关系，R2值为0.998 8；该方法的特异性较好，BLASTN比对，检测引物对仅能匹配滑液支原体的特有基因VY93_RS02885，结果显示，该方法能特异性地检测滑液支原体，与其他常见的引起禽关节炎的细菌/支原体性病原无交叉反应；该方法的敏感性较好，100%阳性检测的靶标基因最小拷贝数为10；该方法的重复性较好，对3个不同浓度的模板进行组间和组内重复检测，变异系数≤1.172 7%。此外，该方法对临床样品的检出率与传统检测方法相符。因此，本研究建立的针对滑液支原体特有基因的SYBR green qPCR检测方法具有用时短、特异性高、准确性高、敏感性高、可重复性强的优点，可为滑液支原体的快速诊断和流行病学调查提供技术手段。
Abstract:
-

参考文献/References:

［1］Dufour-Gesbert F，Dheilly A，Marois C，et al. Epidemiological study on Mycoplasma synoviae infection in layers［J］. Veterinary Microbiology，2006，114(1/2)：148-154.
［2］Kursa O，Pakua A，Tomczyk G，et al. Eggshell apex abnormalities caused by two different Mycoplasma synoviae genotypes and evaluation of eggshell anomalies by full-field optical coherence tomography［J］. BMC Veterinary Research，2019，15(1)：1.
［3］Oh K，Lee S，Seo J，et al. Rapid serodiagnosis with the use of surface plasmon resonance imaging for the detection of antibodies against major surface protein A of Mycoplasma synoviae in chickens［J］. Canadian Journal of Veterinary Research-Revue Canadienne de Recherche Veterinaire，2010，74(1)：71-74.
［4］Kleven S H. Control of avian mycoplasma infections in commercial poultry［J］. Avian Diseases，2008，52(3)：367-374.
［5］Tajadini M，Panjehpour M，Javanmard S H. Comparison of SYBR green and TaqMan methods in quantitative real-time polymerase chain reaction analysis of four adenosine receptor subtypes［J］. Advanced Biomedical Research，2014，3：85.
［6］Gomes C M，Cesetti M V，de Paula N A，et al. Field validation of SYBR green- and TaqMan-Based Real-Time PCR using biopsy and swab samples to diagnose American tegumentary leishmaniasis in an area where Leishmania (Viannia) braziliensis is endemic［J］. Journal of Clinical Microbiology，2017，55(2)：526-534.
［7］Diene S M，Bertelli C，Pillonel T，et al. Comparative genomics of Neisseria meningitidis strains：new targets for molecular diagnostics［J］. Clinical Microbiology and Infection，2016，22(6)：1-7.
［8］Mekkes D R，Feberwee A. Real-time polymerase chain reaction for the qualitative and quantitative detection of Mycoplasma gallisepticum［J］. Avian Pathology，2005，34(4)：348-354.
［9］Huang Y，Niu B，Gao Y，et al. CD-HIT suite：a web server for clustering and comparing biological sequences［J］. Bioinformatics，2010，26(5)：680-682.
［10］Ghanem M，El-Gazzar M. Development of Mycoplasma synoviae (MS) core genome multilocus sequence typing (cgMLST) scheme［J］. Veterinary Microbiology，2018，218：84-89.
［11］Xu B，Liu R，Tao L，et al. Complete genome sequencing of Mycoplasma synoviae strain HN01，isolated from chicken in Henan province，China［J］. Microbiology Resource Announcements，2020，9(6)：e01419-e01480.
［12］Jarquin R，Schultz J，Hanning I，et al. Development of a real-time polymerase chain reaction assay for the simultaneous detection of Mycoplasma gallisepticum and Mycoplasma synoviae under industry conditions［J］. Avian Diseases，2009，53(1)：73-77.
［13］Lauerman L H，Hoerr F J，Sharpton A R，et al. Development and application of a polymerase chain reaction assay for Mycoplasma synoviae［J］. Avian Diseases，1993，37(3)：829-834.
［14］García M，Jackwood M W，Head M，et al. Use of species-specific oligonucleotide probes to detect Mycoplasma gallisepticum，M. synoviae，and M. iowae PCR amplification products［J］. Journal of Veterinary Diagnostic Investigation，1996，8(1)：56-63.
［15］Ramírez A S，Naylor C J，Hammond P P，et al. Development and evaluation of a diagnostic PCR for Mycoplasma synoviae using primers located in the intergenic spacer region and the 23S rRNA gene［J］. Veterinary Microbiology，2006，118(1/2)：76-82.
［16］Fraga A P，de Vargas T，Ikuta N，et al. A multiplex real-time PCR for detection of Mycoplasma gallisepticum and Mycoplasma synoviae in clinical samples from Brazilian commercial poultry flocks［J］. Brazilian Journal of Microbiology，2013，44(2)：505-510.
［17］Sprygin A V，Andreychuk D B，Kolotilov A N，et al. Development of a duplex real-time TaqMan PCR assay with an internal control for the detection of Mycoplasma gallisepticum and Mycoplasma synoviae in clinical samples from commercial and backyard poultry［J］. Avian Pathology，2010，39(2)：99-109.
［18］Shahid M，Markham P F，Marenda M S，et al. High-resolution melting-curve analysis of obg gene to differentiate the temperature-sensitive Mycoplasma synoviae vaccine strain MS-H from non-temperature-sensitive strains［J］. PLoS One，2014，9(3)：e92215.
［19］Zhu L，Konsak B M，Olaogun O M，et al. Identification of a new genetic marker in Mycoplasma synoviae vaccine strain MS-H and development of a strategy using polymerase chain reaction and high-resolution melting curve analysis for differentiating MS-H from field strains［J］. Veterinary Microbiology，2017，210：49-55.
［20］Dijkman R，Feberwee A，Landman W. Development，validation and field evaluation of a quantitative real-time PCR able to differentiate between field Mycoplasma synoviae and the MS-H-live vaccine strain［J］. Avian Pathology ：2017，46(4)：403-415.
［21］Kreizinger Z，Sulyok K M，Grózner D，et al. Development of mismatch amplification mutation assays for the differentiation of MS1 vaccine strain from wild-type Mycoplasma synoviae and MS-H vaccine strains［J］. PLoS One，2017，12(4)：e0175969.
［22］黄莉，谢芝勋，王盛，等. 鸡滑液囊支原体和禽呼肠病毒的实时荧光定量PCR快速检测［J］. 动物医学进展，2018，39(2)：1-6.

相似文献/References:

备注/Memo

备注/Memo:
收稿日期：2021-03-08
基金项目：国家重点研发计划(编号：2017YFD0500706)；国家自然科学基金(编号：32002291)
作者简介：徐彬(1987—)，男，河北廊坊人，博士，助理研究员，主要从事动物传染病防治研究。E-mail：xubin9999@foxmail.com。
通信作者：张小飞，博士，研究员，主要从事动物传染病防治研究。E-mail：xiaofei0804@sina.com。
更新日期/Last Update: 2021-10-05