Collins D M, Stephens D M, de Lisle G W
Central Animal Health Laboratory, Wallaceville Animal Research Centre, Upper Hutt, New Zealand.
Vet Microbiol. 1993 Sep;36(3-4):289-99. doi: 10.1016/0378-1135(93)90095-o.
A polymerase chain reaction (PCR) test for M. paratuberculosis was developed based on a 218 bp segment of a DNA insertion sequence, IS900, that is specific for this organism. The method involved two consecutive amplification reactions, with the second set of primers being nested inside the first set. The method reliably detected 50 organisms/g faeces. This PCR test was applied to 32 bovine faecal specimens containing high, moderate or low numbers of M. paratuberculosis organisms as determined by culture. The PCR test detected all specimens containing > or = 1600 colony forming units (cfu)/g faeces, six of ten specimens with 160-480 cfu/g faeces but only two of 13 specimens containing < or = 112 cfu/g faeces. The sensitivity of this test was better than that of a commercial PCR test which was carried out on the same faecal specimens.
基于副结核分枝杆菌DNA插入序列IS900的一段218 bp片段，开发了一种用于检测副结核分枝杆菌的聚合酶链反应（PCR）试验，该序列对该菌具有特异性。该方法包括两个连续的扩增反应，第二组引物嵌套在第一组引物内部。该方法能可靠地检测出每克粪便中50个菌体。将此PCR试验应用于32份牛粪便标本，这些标本经培养确定含有数量高、中或低的副结核分枝杆菌菌体。PCR试验检测出所有每克粪便中含有≥1600菌落形成单位（cfu）的标本，10份每克粪便中含有160 - 480 cfu的标本中的6份，但13份每克粪便中含有≤112 cfu的标本中仅检测出2份。该试验的灵敏度优于对相同粪便标本进行的商业PCR试验。
