流感嗜血杆菌耐药菌株基因分型的初步研究.docVIP

流感嗜血杆菌耐药菌株基因分型的初步研究林翀 苏应仙(海南省农垦总医院检验科) 摘要：目的 建立随机扩增DNA多态性(RAPD)反应体系对海口市流感嗜血杆菌(Hi)耐药菌株进行基因分型。方法 对临床分离的62株Hi进行药物敏感试验。同时采用RAPD技术对耐药的菌株进行基因分型，并对分型结果进行聚类分析。结果62株Hi中有23株耐药，其中7株(第10~16菌株)对氨苄西林、氯霉素、四环素、复方新诺明耐药，2株(第17~18菌株)对四环素、复方新诺明耐药，1株(第19菌株)对复方新诺明、阿奇霉素耐药，9株(第1~9菌株)对氨苄西林耐药和4株(第20~23株)对阿奇霉素耐药。采用引物(5-ACGTATCTGC-3)构建的RAPD-PCR反应体系可以扩增出质量较好的带谱所构建的亲缘关系聚类图中，第1~9菌株聚成一，第10~18菌株聚成一，第19菌株聚成一，第20~23菌株聚成一。结论：RAPD-PCR反应体系可以较好地进行Hi耐药菌株的基因分型。 关键词：流感嗜血杆菌；随机扩增DNA多态性；基因分型；耐药性 Application of randomly amplified polymorphic DNA technique in homological analysis of Haemophilus influenza(Laboratory, Nongken Chief Board Hospital of Hainan Province,HaiKou 570311,China ) Abstract: The aim of this study is to analyze the homological analysis of Haemophilus influenza in Haikou by randomly amplified polymorphic DNA (RAPD) technique. Methods: The RAPD technique was used in genotype of 62 Haemophilus influenza strains isolated from clinical samples, and cluster analysis was performed in RAPD results. Results: There were 23 drug resistant strains in 62 Hi strains, consisting of 7 strains (No.10-16) resistant to ampicillin, chloromycetin, tetracycline, sulfamethoxazole compound, 2 strains (No.17-18) resistant to tetracycline, sulfamethoxazole compound, 1 strain (No.19) resistant to sulfamethoxazole compound, azithromycin, 9 strains (No.1-9) resistant to ampicillin and 4 strains (No.20-23) resistant to azithromycin. The maps with good quality were obtained with RAPD-PCR systems using random primer (5-ACGTATCTGC-3). The relationship dendrogram mapped with random primer showed the 23 drug resistant strains were divided into 4 clusters, including a cluster with No.1-9 strain, a cluster with No.10-18 strain, a cluster with No.19 strain and a cluster with No.20-23 strain. Conclusion: Genotype identification for Haemophilus influenza strains using RAPD method is significant for determining the genetic homology. Key words: Haemophilus influenza; Randomly amplified polymorphic DNA; Genotype; Drug resistance 流感嗜血杆