摘要
[目的]探讨PCR-反向膜杂交技术鉴定疑似关节结核临床标本中分枝杆菌菌种的应用价值。[方法]应用PCR-反向膜杂交法和培养菌种鉴定法对35例疑似关节结核和35例非关节结核临床标本行分枝杆菌菌种鉴定,应用PCR-直接测序法验证PCR-反向膜杂交法阳性鉴定结果准确性。[结果]培养菌种鉴定法在疑似关节结核组培养鉴定出8例结核分枝杆菌,1例牛结核分枝杆菌,非关节结核组培养结果均为阴性;灵敏度25.7%,特异度100%。PCR-反向膜杂交法在疑似关节结核组鉴定出18例结核分枝杆菌复合群,1例戈登分枝杆菌,1例浅黄分枝杆菌,与PCR-直接测序结果一致;非关节结核组中1例结核分枝杆菌复合群阳性,PCR-直接测序未扩增出核苷酸序列;灵敏度57.1%,特异度97.1%。两种菌种鉴定法检测阳性率差异有统计学意义(x^2=11.28,P=0.01)。[结论]PCR-反向膜杂交技术可快速将疑似关节结核中分枝杆菌鉴定至种,指导诊断。
[Objective] To evaluate the application value of PCR-reverse cross-blot hybridization method in the rapid identification of mycobacterium species in suspicious joint tuberculosis. [Methods] PCR-reverse cross-blot hybridization and culture combined with strain identification were used to identify the species of mycobacteria in 35 patients with suspicious joint tuberculosis,compared with clinical specimens of 35 cases of non-joint tuberculosis,PCR-direct sequencing was also used to verify the accuracy of positive results identified by PCR-reverse cross-blot hybridization method. [Results] Culture and strain identification found 8 cases of mycobacterium tuberculosis and 1 case of mycobacterium bovis in the suspicious joint tuberculosis group,whereas all of the results were negative in the non-joint tuberculosis group; associated with sensitivity of25. 7% and specificity of 100%. By contrast,PCR-reverse cross-blot hybridization identified 18 cases of mycobacterium tuberculosis complex,1 case of mycobacterium gordonae and 1 case of mycobacterium gilvum in the suspicious joint tuberculosis group,while only a case of mycobacterium tuberculosis complex in the non-joint tuberculosis group,with sensitivity of 57. 1%and specificity of 97. 1%. The results of PCR-reverse cross-blot hybridization were consistent with those of PCR-direct sequencing which could not amplify nucleotide sequences. There was a significant difference of positive ratio in the results between PCR-reverse cross-blot hybridization and culture combined with strain identification（ x^2= 11. 28,P = 0. 01）. [Conclusion] PCR-reverse cross-blot hybridization,with advantage of quick identification of mycobacterium tuberculosis into species,should be used to assist accurately making diagnosis.
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
2016年第7期653-656,共4页
Orthopedic Journal of China
基金
首都临床特色应用研究项目(编号:Z1211070010112094)
国家重大传染病防治科技重大专项项目(编号:2013ZX10003003005)
军队后勤"十二五"科研课题重点项目(编号:11MS011)
关键词
关节结核
PCR-反向膜杂交
分枝杆菌
菌种鉴定
joint tuberculosis
PCR-reverse cross-blot hybridization
mycobacterium
strain identification
