CAJ | 학술논문

目的：避免霍乱弧菌检测假阳性，提高检测正确率。方法随机选取2013年1～7月，全国各省市 CDC 送往中国CDC 霍乱初筛阳性菌株14株；用 LB 营养琼脂培养12 h，挑取单菌落进行霍乱弧菌血清凝集，同时水煮模板法提取菌株的DNA。针对弧菌属16SrDNA 序列设计引物，进行弧菌16SrDNA PCR 检测，电泳观察16SrDNA 产物，将16SrDNA 阳性产物送测序公司测序，测序结果在 NCBI 网站上进行 Blast 比对，分析比较血清凝集和 Blast 比对结果。结果共选取14株菌进行实验，血清凝集阳性12株，2株未凝。经弧菌16SrDNA 扩增，电泳观察14株菌均扩增出相应的片段，说明所选菌株均为弧菌属。将14株菌的16SrDNA 阳性产物测序，并将测序结果进行 Blast 比对：2株血清未凝集菌均是哈维氏弧菌；12株血清凝集阳性的菌中，1株是需钠弧菌，其余11株是霍乱弧菌。结论血清凝集和基因测序联合检测群霍乱弧菌，可避免霍乱弧菌检测假阳性，提高检测正确率。
목적：피면곽란호균검측가양성，제고검측정학솔。방법수궤선취2013년1～7월，전국각성시 CDC 송왕중국CDC 곽란초사양성균주14주；용 LB 영양경지배양12 h，도취단균락진행곽란호균혈청응집，동시수자모판법제취균주적DNA。침대호균속16SrDNA 서렬설계인물，진행호균16SrDNA PCR 검측，전영관찰16SrDNA 산물，장16SrDNA 양성산물송측서공사측서，측서결과재 NCBI 망참상진행 Blast 비대，분석비교혈청응집화 Blast 비대결과。결과공선취14주균진행실험，혈청응집양성12주，2주미응。경호균16SrDNA 확증，전영관찰14주균균확증출상응적편단，설명소선균주균위호균속。장14주균적16SrDNA 양성산물측서，병장측서결과진행 Blast 비대：2주혈청미응집균균시합유씨호균；12주혈청응집양성적균중，1주시수납호균，기여11주시곽란호균。결론혈청응집화기인측서연합검측군곽란호균，가피면곽란호균검측가양성，제고검측정학솔。
Objective To avoid false positive detection ofVibrio cholerae and improve the detection correct rate.Methods 1~7 months of 2013 were randomly selected,the national various provinces and cities CDC to China cholera CDC positive screening 14 strains.LB nutrient agar 12 hours,take single colony to Vibrio cholera serum agglutination,extraction of strain DNA at the same time boiled template method.For Vibrio 16SrDNA sequence and design primers for PCR detection of Vib-rio,16SrDNA,electrophoresis were used to observe the 16SrDNA products,16SrDNA positive products sent to sequencing company sequencing,sequencing results were Blast comparison on the NCBI website for the analysis and comparison of ser-um agglutination and Blast alignment.Results 12 strains was positive for agglutination and 2 strains of non agglutination in 14 strains.The Vibrio 16SrDNA amplification,electrophoresis were used to observe the 14 isolates that were amplified frag-ment corresponding,that the selected strains were vibrio.The 16SrDNA positive products were 14 strains,and the sequen-cing the Blast results:2 strains of bacteria were Vibrio harveyi for non agglutination,in 12 positive strains of serum aggluti-nation;1 strains was Vibrio natriegen and 11 strains wereVibrio cholerae .Conclusion Detection ofVibrio cholerae cholerae combined with serum agglutination and gene sequencing can avoid false positive result ofVibrio cholerae ,and improve cor-rect rate of the detection.