Lau Q C, Chow V T, Poh C L
Department of Microbiology, Faculty of Medicine, National University of Singapore, Republic of Singapore.
Genitourin Med. 1995 Dec;71(6):363-6. doi: 10.1136/sti.71.6.363.
To employ polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis for the rapid differentiation of Neisseria gonorrhoeae protein IB (PIB) isolates and to compare its usefulness with the widely accepted auxotype/serovar classification scheme.
The outer membrane protein IB genes of 47 gonococcal isolates belonging to 10 different serovars were amplified by PCR. The approximately 1 kb DNA products were then digested separately with restriction enzymes CfoI and MspA1I, and electrophoresed on agarose gels.
Cleavage of PIB genes by MspA1I and CfoI differentiated all the N gonorrhoeae strains into five and six PCR-RFLP profiles, respectively. PCR-RFLP was more discriminatory than auxotyping, which classifies the strains into only two auxotypes. Some strains belonging to common serovars could be further differentiated. A combination of PCR-RFLP analysis, auxotyping and serotyping further increased the discrimination of the strains into 34 subtypes. The PCR-RFLP method was easy to perform, reliable, reproducible, and consistent with published nucleotide sequence data.
The PCR-RFLP method can augment auxotyping and serotyping or be used as a preliminary screening tool to differentiate N gonorrhoeae strains in areas where serotyping reagents are not easily available.
采用聚合酶链反应（PCR）和限制性片段长度多态性（RFLP）分析快速鉴别淋病奈瑟菌蛋白IB（PIB）分离株，并将其与广泛接受的营养型/血清型分类方案的实用性进行比较。
通过PCR扩增47株属于10种不同血清型的淋球菌分离株的外膜蛋白IB基因。然后分别用限制性内切酶CfoI和MspA1I消化约1 kb的DNA产物，并在琼脂糖凝胶上进行电泳。
MspA1I和CfoI对PIB基因的切割分别将所有淋病奈瑟菌菌株分为5种和6种PCR-RFLP图谱。PCR-RFLP比营养型分型更具鉴别力，营养型分型只能将菌株分为两种营养型。一些属于常见血清型的菌株可以进一步区分。PCR-RFLP分析、营养型分型和血清型分型相结合进一步将菌株的鉴别增加到34个亚型。PCR-RFLP方法操作简便、可靠、可重复，且与已发表的核苷酸序列数据一致。
PCR-RFLP方法可增强营养型分型和血清型分型，或用作在血清型分型试剂不易获得的地区鉴别淋病奈瑟菌菌株的初步筛选工具。
