Rasmussen H N, Rasmussen O F, Andersen J K, Olsen J E
Biotechnological Institute, Lyngby, Denmark.
Mol Cell Probes. 1994 Apr;8(2):99-108. doi: 10.1006/mcpr.1994.1014.
A pair of polymerase chain reaction (PCR) primers, YC1 and YC2, selected from the sequence of the invasin locus (inv) of Y. enterocolitica, has been evaluated for specific detection of pathogenic Y. enterocolitica by PCR. The primers were hybridized at high stringency conditions to DNA from 65 pathogenic Y. enterocolitica, 16 non-pathogenic Y. enterocolitica, 18 other Yersinia and 124 non-Yersinia strains. YC2 hybridized to the pathogenic Y. enterocolitica only, while YC1 hybridized weakly to nine non-Yersinia as well. In a PCR with annealing at 64 degrees C all Y. enterocolitica, pathogenic and non-pathogenic, were positive. However, DNA from 60 non-Y. enterocolitica was amplified. With annealing at 72 degrees C, 10 non-pathogenic Y. enterocolitica and 41 non-Y. enterocolitica were positive. When a two-step PCR assay with annealing at 72 degrees C, hot-start and 1% dimethylsulphoxide (DMSO) were used, only DNA from the pathogenic Y. enterocolitica were amplified. The limit of detection was shown for four different strains to be less than 10 cells per PCR tube.
一对从小肠结肠炎耶尔森氏菌侵袭素基因座（inv）序列中选出的聚合酶链反应（PCR）引物YC1和YC2，已通过PCR用于致病性小肠结肠炎耶尔森氏菌的特异性检测评估。这些引物在高严格条件下与来自65株致病性小肠结肠炎耶尔森氏菌、16株非致病性小肠结肠炎耶尔森氏菌、18株其他耶尔森氏菌和124株非耶尔森氏菌菌株的DNA杂交。YC2仅与致病性小肠结肠炎耶尔森氏菌杂交，而YC1也与9株非耶尔森氏菌弱杂交。在退火温度为64℃的PCR中，所有致病性和非致病性小肠结肠炎耶尔森氏菌均呈阳性。然而，60株非小肠结肠炎耶尔森氏菌的DNA被扩增。在退火温度为72℃时，10株非致病性小肠结肠炎耶尔森氏菌和41株非小肠结肠炎耶尔森氏菌呈阳性。当使用退火温度为72℃、热启动和1%二甲基亚砜（DMSO）的两步PCR检测法时，仅致病性小肠结肠炎耶尔森氏菌的DNA被扩增。四种不同菌株的检测限显示为每个PCR管少于10个细胞。
