利福平耐药实时荧光定量核酸扩增检测技术对脊柱结核不同类型标本检测的研究

A retrospective study on the role of automated real - time nucleic acid amplification technology for rapid and simultaneous detection of mycobacterium tuberculosis and rifampicin resistance in testing different types of tissue specimens of spinal tuberculosis

目的比较应用利福平耐药实时荧光定量核酸扩增检测技术（XpertMTB／RIF）对脊柱结核不同类型组织标本检测的差异。方法收集在天津市海河医院骨科接受病灶清除手术治疗诊断明确的脊柱结核患者223例，收集脓液、肉芽组织及干酪样坏死组织，分别进行结核分枝杆菌培养和XpertMTB／RIF检测，计算阳性率，并以细菌培养与表型药敏试验为金标准，计算XpertMTB／RIF进行细菌和利福平耐药基因rpoB突变检测的敏感度、特异度。结果223例脊柱结核患者多标本联合XpertMTB／RIF检测阳性率高于细菌培养阳性率，差异有统计学意义[（68．61±7．35）％比较（44．39±6．51）％，P=0．000]。两种细菌检测方法中，都是脓液阳性率最高，肉芽组织其次，干酪样坏死组织最低，差异有统计学意义[细菌培养：（44．44±7．08）％比（43．86±7．44）％比（30．72±7．3l％），P=0．017；XpertMTB／RIF检测：（73．02±6．33）％比（63．74±7．21）％比（39．87±7．76％），P=0．000]。3种标本XpertMTB／RIF检测阳性率都高于细菌培养（脓液、肉芽组织：P=0．000，差异有统计学意义；干酪样坏死组织：P=0．131，差异无统计学意义）。XpertMTB／RIF检测结核杆菌的敏感度达（96．97±3．38）％，rpoB突变检测的敏感度（97．44％）和特异度（94．74％）也较高。结论脊柱结核患者应尽量获取多种标本，尤其脓液标本，进行XpertMTB／RIF检测，可简便、快速、准确地诊断结核感染及评估其耐药性，有助于开展有效治疗。

Objective To compare the role of automated real - time nucleic acid amplification technology for rapid and simultaneous detection of mycobacterium tuberculosis and rifampicin resistance （Xpert MTB/RIF） in testing different types of tissue specimens of spinal tuberculosis. Methods The pus, granulation tissue and caseous necrotic tissue of 223 patients who were diagnosed as having spinal tu- berculosis （TB） and received curettage at the Department of Orthopedics of Tianjin Haihe Hospital were collected for bacterial culture and Xpert MTB/RIF assay. Also, the bacterial culture and phenotypic drug- sensitivity test were adopted as the golden standards to calculate the sensitivity and specificity of Xpert MTB/RIF assay. Results The positive rate of Xpert MTB/RIF assays on spinal TB patients＇ tissue specimens was higher than that of bacterial culture [ （ 68.61± 7.35 ） % vs. （ 44. 39±6. 51 ） % , P = 0. 000 ]. The positive rate of pus was the highest, followed by granulation tissue, and the caseous necrotic tissue was the lowest by both methods with statistically significant difference [ bacterial culture： （44.44 ± 7.08）% 〉 （43.86±7.44）% 〉 （30.72±7.31%）, P=0.017; Xpert MTB/RIF assay： （73.02±6. 33 ） % 〉 （63.74 ±7.21 ） % 〉 （39. 87± 7.76% ）, P = 0. 000 ]. The positive rate of Xpert MTB/RIF assay was higher than that of bacterial culture in all three types of specimens （ pus and granulation tissue ： P = O. 000, with statistically significant difference; caseous necrotic tissue： P = 0. 131, without statistically significant difference）. The sensitivity of Xpert MTB/RIF assay on mycobacterium tuberculosis was up to （96. 97 ± 3.38） % while the sensitivity （97.44%） and specificity （94. 74% ） of drug - sensitivity test al- so remained relatively high. Conclusion For efficient and accurate diagnosis of TB and drug - resistance test and timely provision of effective treatment, multiple specimens, especially the pus of spinal TB pa- tients, should be collected for Xpert MTB/RIF assays.