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為了解我國(guó)豬肺炎支原體種群結(jié)構(gòu)和進(jìn)化關(guān)系,2018—2019年采集山東、廣西、河南、江西、云南、天津、湖南等地豬組織樣品1 242份,進(jìn)行豬肺炎支原體熒光定量PCR和巢式PCR檢測(cè),并對(duì)測(cè)序確認(rèn)的87份陽性樣品,采用adk、rpoB、tpiA 3個(gè)管家基因作為分型參考進(jìn)行多位點(diǎn)序列分型(MLST),并對(duì)3個(gè)管家基因均擴(kuò)增成功的陽性樣品進(jìn)行p146基因分型,分析我國(guó)豬肺炎支原體種群結(jié)構(gòu),并結(jié)合pubmlst數(shù)據(jù)庫(kù)公布的151個(gè)序列類型(ST)進(jìn)行eBURST分析。結(jié)果顯示:有24份陽性樣品的3個(gè)管家基因全部擴(kuò)增且測(cè)序成功,共分為10個(gè)ST,均為國(guó)內(nèi)外首次報(bào)道;10個(gè)ST分為3個(gè)克隆復(fù)合體(CC)和4個(gè)獨(dú)特型,其中ST128占41.7%(10/24),樣品來自江蘇、河南和廣西,為本次檢測(cè)的主要流行型。結(jié)果表明,我國(guó)豬肺炎支原體分布較廣,具有一定的基因型獨(dú)特性。本研究為我國(guó)豬肺炎支原體的流行病學(xué)數(shù)據(jù)庫(kù)研究提供了新資料,有助于掌握國(guó)內(nèi)豬肺炎支原體基因型分布情況,進(jìn)一步完善了豬肺炎支原體流行病學(xué)數(shù)據(jù)庫(kù)。
Multi-locus Sequence Typing of Mycoplasma hyopneumoniae in Some Regions of China during 2018 to 2019
In order to investigate the genetic characteristics and evolutionary relationship of Mycoplasma hyopnumoniae(M. hyopnumoniae)in China,1 242 swine tissues were collected during 2018 to 2019 from Shandong,Guangxi,Henan,Jiangxi,Yunnan,Tianjin and Hunan for detection of M. hyopnumoniae by real-time PCR and nested PCR,and 87 positive samples were confirmed. DNA was extracted from positive samples and classified according to p146 gene and multi-locus sequence typing(MLST)with adoption of 3 housekeeping genes including adk,rpoB and tpiA,then the population structure of M. hyopnumoniae in China was analyzed,and eBURST analysis was carried out in combination of 151 STs published in pubmlst database. The results showed that all the three housekeeping genes of 24 positive samples were successfully amplified and sequenced,which were divided into 10 STs and firstly reported in the world,involving three clone complexes(CCs)and four singletons,in which,ST128 was prevalent and accounted for 41.7%(10/24)and the samples were collected from Jiangsu,Henan and Guangxi. It was concluded that M. hyopnumoniae spread widely with unique genotypes to some extent. Therefore,some new data was provided for the study on epidemiological database of M. hyopnumoniae,which contributed to the identification of its distribution and further enriched relevant database.
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國(guó)家獸藥產(chǎn)業(yè)技術(shù)創(chuàng)新聯(lián)盟 National veterinary drug industry technology innovation alliance |
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