荧光定量PCR检测结核/非结核分枝杆菌在肉芽肿病理诊断中的应用

Application of quantitative PCR detection of TB/non-tuberculous mycobacteria in pathological diagnosis of granulomatous lesions

目的探讨荧光定量PCR技术检测肉芽肿病变中结核/非结核性分枝杆菌的可靠性和敏感性。方法采用特殊染色(抗酸、金胺O染色)及荧光定量PCR技术对200例肉芽肿病变中的分枝杆菌进行检测。结果 200例肉芽肿病变中62例抗酸染色(31%)阳性,81例金胺O染色(40.5%)阳性,103例荧光定量PCR检测(51.50%)分枝杆菌阳性,其中100例(50.00%)检出结核杆菌,3例(1.5%)检出非结核分枝杆菌,3种方法检测出的阳性率差异有统计学意义(P<0.05)。典型结核组的抗酸染色阳性率(51.5%)和金胺O染色阳性率(65.7%)及荧光定量PCR检测结核杆菌阳性率(75.8%)皆高于非典型结核组(10.9%、15.8%及24.8%),差异均具有统计学意义(P<0.05)。97例荧光定量PCR检测阴性的标本中15例(15.5%)呈抗酸染色和(或)金胺O染色阳性,82例标本(84.5%)荧光定量PCR检测、金胺O染色和抗酸染色检测均阴性。82例特殊染色和荧光定量PCR检测结核分枝杆菌均阴性标本进行荧光定量PCR重复检测,结核分枝杆菌阳性率典型结核组为35.5%,疑似结核组为3.9%;200例标本荧光定量PCR重复检测后总阳性率从51.5%增至56.5%。结论采用荧光定量PCR检测病变组织中的分枝杆菌,对结核/非结核分枝杆菌分类有效,阳性率高于特殊染色(抗酸、金胺O染色),为临床诊治提供可靠依据。

Purpose To detect TB/non-tuberculous mycobacteria in granulomatous lesions by using fluorescence quantitative PCR method, and to analyze the reliability and sensitivity of the detection method. Methods Special stain （acid-fast auramine O staining） and quantitative PCR method were used for detection of myeobactefium and diagnosis of 200 cases of granulomatous lesions. Results In 200 cases of granulomatous lesions detected, 62 patients （31% ） were positive for acid-fast staining; by auramine O staining 81 cases （40. 5% ） were positive; 103 cases （ 51.50% ） were positive for myeobacterium by quantitative PCR, of which 100 eases （50. 0% ） of mycobacterium tuberculosis were detected, and 3 cases （1.5%） of non-tuberculous mycobaeteria; The positive rate of three methods had statistically significant difference （P 〈 0.05 ）. Typical acid-fast stain positive for tuberculosis group （51.5%） , positive rate （65.7%） of auramine O staining and quantitative PCR Mycobaeterium tuberculosis-positive rate （75.8%） were higher than that of atypical tuberculosis group （ 10. 9%, 15.8% and 24. 8% ）, with statistically significant differences （ P 〈 0.05 ）. In 97 specimens with negative quantitative PCR result, acid-fast staining and/or auramine O was positive in 15 cases （ 15.5% ）, and all quantitative PCR, auramine O and acid-fast staining were negative in 82 cases （84. 5% ）. In 82 cases with negative special staining and PCR results, the duplicate detection of quantitative PCR showed that positive Mycobaeterium tuberculosis rate was 35.5% in typical tuberculosis group, and 3.9% in suspected tuberculosis group; In the 200 cases of the total positive rate after quantitative PCR increased from 51.5% to 56. 5%. Conclusions Myeabacterium quantitative PCR assay can be effective in the classification of TB/nontuberculous myeobacteria, and it is more sensitive than other special staining methods （acid-fast, auramine 0 staining）, which pro- vides a reliable basis for the clinical diagnosis and treatment.