Pan Tzu-Ming, Liu Yi-Ju
Department ofAgricultural Chemistry, National Taiwan University, Taipei, ROC.
J Microbiol Immunol Infect. 2002 Sep;35(3):147-51.
The polymerase chain reaction and the multiplex polymerase chain reaction were developed for detection of Salmonella and for identification of the serotype enteritidis. Three sets of primers were selected from different genomic sequences amplifying a 429 bp fragment specific for the genus Salmonella within a randomly cloned sequence, including a 250 bp fragment within the spv gene, and a 310 bp fragment within the sefA gene specific for Salmonella enteritidis. The polymerase chain reaction and the multiplex polymerase chain reaction were used for detecting S. enteritidis isolated from stool samples during outbreaks of foodborne gastroenteritis between 1992 and 1998 in Taiwan. The sefA gene was detected in all 27 strains of S. enteritidis by this polymerase chain reaction method. Multiplex polymerase chain reaction could detect 3 genes in all strains, but could not detect the spv gene in 2 strains. The sensitivity of the polymerase chain reaction and the multiplex polymerase chain reaction were 10(4) and 10(5) cells/mL, respectively. In double polymerase chain reaction, the sensitivity increased to 100 cells/mL. These data indicate that the specificity and sensitivity of the polymerase chain reaction and the multiplex polymerase chain reaction make them potentially valuable tools for diagnosis of S. enteritidis infection and that they may be used for the identification of S. enteritidis responsible for sporadic enteritis cases.
聚合酶链反应（PCR）和多重聚合酶链反应被用于检测沙门氏菌以及鉴定肠炎血清型。从不同基因组序列中选取了三组引物，在一个随机克隆的序列中扩增出一段对沙门氏菌属特异的429 bp片段，包括spv基因内的一段250 bp片段以及肠炎沙门氏菌特异的sefA基因内的一段310 bp片段。1992年至1998年期间，在台湾食源性肠胃炎暴发期间，使用聚合酶链反应和多重聚合酶链反应检测从粪便样本中分离出的肠炎沙门氏菌。通过这种聚合酶链反应方法，在所有27株肠炎沙门氏菌中均检测到了sefA基因。多重聚合酶链反应可以在所有菌株中检测到3个基因，但有2株菌株未能检测到spv基因。聚合酶链反应和多重聚合酶链反应的灵敏度分别为10(4)和10(5)个细胞/mL。在双重聚合酶链反应中，灵敏度提高到了100个细胞/mL。这些数据表明，聚合酶链反应和多重聚合酶链反应的特异性和灵敏度使其成为诊断肠炎沙门氏菌感染的潜在有价值工具，并且它们可用于鉴定导致散发性肠炎病例的肠炎沙门氏菌。
