【摘要】：目的用PCR結(jié)合酶切-序列比對(duì)法對(duì)B群腦膜炎奈瑟菌菌株進(jìn)行鑒定。方法用玻片凝集法對(duì)不同來源的15株B群腦膜炎奈瑟菌菌株進(jìn)行初步檢定,再用PCR結(jié)合酶切-序列比對(duì)法對(duì)上述15株菌株進(jìn)行進(jìn)一步鑒定,即用PCR結(jié)合酶切法擴(kuò)增菌株的唾液酸轉(zhuǎn)移酶sia D基因并對(duì)PCR產(chǎn)物進(jìn)行酶切后,用BLAST軟件將PCR產(chǎn)物測(cè)序結(jié)果與Gene Bank中原始sia D序列比對(duì)。結(jié)果 15株菌株玻片凝集結(jié)果均為陽性;15株菌株的PCR產(chǎn)物片段大小均為460 bp;TaqⅠ酶切后,13株菌株的酶切產(chǎn)物片段大小仍為460 bp,其PCR產(chǎn)物測(cè)序比對(duì)結(jié)果與B群腦膜炎奈瑟菌原始sia D序列同源性均達(dá)到99%;其余2株酶切產(chǎn)物片段大小約200 bp,與C群腦膜炎奈瑟菌sia D原始基因序列同源性分別為98%和99%。結(jié)論 15株菌株經(jīng)PCR結(jié)合酶切-序列比對(duì)法鑒定,13株為B群腦膜炎奈瑟菌菌株,2株為C群腦膜炎奈瑟菌菌株;該方法可準(zhǔn)確鑒定B群腦膜炎奈瑟菌菌株。
[Abstract]:Objective to identify Neisseria meningitidis group B by PCR and enzyme-sequence alignment. Methods 15 strains of Neisseria meningitidis from different sources were preliminarily identified by slide agglutination method. The 15 strains were further identified by PCR combined with enzyme-sequence alignment method. The sialic acid transferase sia D gene of the strain was amplified by PCR binding enzyme digestion method and the PCR product was digested. The results of PCR product sequencing were compared with the original sia D sequence in Gene Bank by BLAST software. Results the results of slide agglutination of 15 strains were all positive. The size of PCR products of 15 strains were all 460bpTQ 鈪，

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