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目的用PCR结合酶切-序列比对法对B群脑膜炎奈瑟菌菌株进行鉴定。方法用玻片凝集法对不同来源的15株B群脑膜炎奈瑟菌菌株进行初步检定,再用PCR结合酶切-序列比对法对上述15株菌株进行进一步鉴定,即用PCR结合酶切法扩增菌株的唾液酸转移酶sia D基因并对PCR产物进行酶切后,用BLAST软件将PCR产物测序结果与Gene Bank中原始sia D序列比对。结果 15株菌株玻片凝集结果均为阳性;15株菌株的PCR产物片段大小均为460 bp;TaqⅠ酶切后,13株菌株的酶切产物片段大小仍为460 bp,其PCR产物测序比对结果与B群脑膜炎奈瑟菌原始sia D序列同源性均达到99%;其余2株酶切产物片段大小约200 bp,与C群脑膜炎奈瑟菌sia D原始基因序列同源性分别为98%和99%。结论 15株菌株经PCR结合酶切-序列比对法鉴定,13株为B群脑膜炎奈瑟菌菌株,2株为C群脑膜炎奈瑟菌菌株;该方法可准确鉴定B群脑膜炎奈瑟菌菌株。
Objective To identify Neisseria meningitidis strain B by PCR combined with enzyme digestion-sequence alignment. Methods Fifteen strains of Neisseria meningitidis group B from different sources were preliminarily tested by slide agglutination method. The 15 isolates were further identified by PCR and enzyme digestion-sequence alignment. After amplifying the sial D gene of sialic acid transferase strain and digesting the PCR product, the sequencing results of the PCR product were compared with the original sia D sequence in Gene Bank by BLAST software. Results The results of slide agglutination were positive in all 15 strains. The size of PCR products of the 15 isolates was 460 bp. After digested with TaqⅠ, the sizes of the digested products of the 13 isolates were still 460 bp. The PCR products were sequenced The results were 99% identical to the original sia D sequence of Neisseria meningitidis group B, and the size of the other two fragments was about 200 bp. The sequence homology with the original gene of sia D of N. meningitidis group C 98% and 99%. Conclusion 15 strains were identified by PCR combined with enzyme digestion-sequence alignment, 13 strains were Neisseria meningitidis group B and 2 strains were Neisseria meningitides group C. This method can accurately identify meningitis group B Candida strains.