A Molecular Epidemiological Study on Mycoplasma Pneumonia of Sheep and Goats in Fujian
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摘要: 为了解福建羊支原体性肺炎和山羊传染性胸膜肺炎的分子流行情况，于2014-2016年从福建省6个市采集到疑似羊支原体性肺炎或山羊传染性胸膜肺炎病料61份，用支原体目引物、绵羊肺炎支原体、丝状支原体山羊亚种、山羊支原体山羊肺炎亚种、山羊支原体山羊亚种、精氨酸支原体、莱氏无胆甾原体和无乳支原体的特异引物，通过PCR方法进行检测。挑取部分PCR阳性病料进行支原体分离和纯化，用16S rRNA基因通用引物进行克隆测序。结果显示61份样品中检出绵羊肺炎支原体45份（73.77%），检出丝状支原体山羊亚种4份（6.56%），检出莱氏无胆甾原体4份（6.56%），检出精氨酸支原体2份（3.28%）。该研究结果表明福建省发生的类传染性胸膜肺炎是以绵羊肺炎支原体为主的羊支原体性肺炎，为福建省羊支原体性肺炎的有效防治提供了科学依据。Abstract: The molecular epidemiology on the mycoplasma pneumonia of sheep and the contagious caprine pleuropneumonia of goats in Fujian were investigated. A total of 61 samples from the goats with suspected infection of the diseases were collected from 6 regions in the province during 2014-2016. PCR with the mycoplasmatales primer as well as the specific primers for Mycoplasma ovipneumoniae, Mycoplasma mycoides subsp.capri, Mycoplasma capricolum Subsp. Capripneumonia, Mycoplasma capricolum subsp.capri, Mycoplasma arginin, Acholeplasma laidlawii and Mycoplasma agalactiae were applied for the detection.Part of the PCR positive clinical samples were selected to isolate Mycoplasma for subsequent cloning and sequencing based on their 16S rRNA. The detection rates on M. ovipneumoniae, M. mycoides subsp.capri, A. laidlawii and M. arginine were 73.77%(45/61), 6.56%(4/61), 6.56%(4/61) and 3.28%(2/61), respectively. The results suggested that the suspected contagious caprine pleuropneumonia in the province were mycoplasma pneumonia of sheep and goats caused mainly by M.ovipneumoniae. The finding would lead to effective prevention and treatment measures for the diseases.
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图 1 部分样品支原体目PCR检测结果
注：M为DNA Marker 2000；PC为Y98标准株，NC为阴性对照，09~18、22~24、26~33、35均为病料样品。
Figure 1. PCR detection with mycoplasmatale primers in some samples
图 2 部分样品绵羊肺炎支原体PCR检测结果
注：M为DNA Marker 2000；PC为Y98标准株，NC为阴性对照，07、09~16、18、22~25、27、28、30~33、35、36均为病料样品。
Figure 2. PCR detection of M.ovipneumoniae in some samples
图 3 部分样品丝状支原体山羊肺炎亚种PCR检测结果
注：M为DNA Marker 2000；PC为PG3标准株，NC为阴性对照，29~33为病料样品。
Figure 3. PCR detection of M. mycoides subsp. Capri in some samples
图 4 部分样品莱氏无胆甾原体PCR检测结果
注：M为DNA Marker 2000；10、11、40、47~50均为病料样品。
Figure 4. PCR detection of A. laidlawii in some samples
图 5 部分样品精氨酸支原体PCR检测结果
注：M为DNA Marker 2000；28、29、31、33、35~37均为病料样品。
Figure 5. PCR detection of M. arginini in some samples
表 1 参比序列
Table 1. Reference sequence
支原体种类 缩写名称 基因库
序列号Acholeplasma laidlawiistrain
PG-8AAL PG8 NR_074448.1 Mycoplasma mycoidessubsp.
capri strain PG3Mmc PG3 NR_044772.1 Mycoplasma mycoides mycoides
LC type,Y-goatMmmLC Y-goat U26044.1 Mycoplasma capricolum
capricolumG5Mcc G5 U26041.1 Mycoplasma capricolumsubsp.
capripneumoniae strain F38Mccp F38 NR_037061.1 Mycoplasma ovipneumoniae
strain Y-98Mo Y98 NR_025989.1 Mycoplasma agalactiae strain PG2 Maga PG2 NR_118811.1 Mycoplasma arginini strain G230 Marg G230 NR_041743.1 
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