荧光定量PCR技术在骨关节结核石蜡包埋标本检测中的应用价值

Application value of fluorescence quantitative PCR technology in detection of paraffin-embedded specimens from bone and joint tuberculosis

目的探讨荧光定量聚合酶链式反应（fluorescent quantitative polymerase chain reaction，FQPCR）在骨关节结核石蜡包埋标本病理诊断及鉴别诊断中的应用价值。方法搜集2014年10月至2015年4月首都医科大学附属北京胸科医院病理科诊断形态学符合结核的93例患者的石蜡包埋标本，12例骨肿瘤患者的石蜡包埋标本。所有标本以FQ-PCR技术检测结核分枝杆菌DNA，以抗酸染色法查找抗酸杆菌。以卡方检验分析实验数据，比较FQ-PCR技术与传统抗酸染色法在骨关节结核诊断中的敏感度和特异度；观察不同发病部位与两种技术检测阳性率之间的相关性；以PCR-反向点杂交法对7例结核分枝杆菌DNA阴性而抗酸染色阳性患者进行非结核分枝杆菌检测。结果所有93例骨关节结核标本中FQPCR技术检测阳性77例，敏感度82．8％；抗酸染色查到抗酸杆菌64例，敏感度68．8％。FQ-PCR技术检测敏感度明显高于抗酸染色法（X2=5．659，P=0．021）。在不同发病部位FQ-PCR技术敏感度均高于抗酸染色法：在脊柱病变FQ-PCR技术和抗酸染色法阳性率分别为84．0％（42／50）和64．0％（32／50）（X2=0．009，P=0．609）；在外周骨关节病变FQ-PCR技术和抗酸染色法阳性率分别为81．4％（35／43）和74．4％（32／43）（X2=12．609，P=0．002）。PCR-反向点杂交法检测确定戈登分枝杆菌1例。结论FQ-PCR技术较抗酸染色法显著提高了结核分枝杆菌阳性检出率，并为进一步检测非结核分枝杆菌做出重要提示，FQ-PCR技术在骨关节结核石蜡包埋标本检测中具有良好的应用价值。

Objective To explore the value of fluorescence quantitative PCR （FQPCR） technique in the diagnosis and differential diagnosis of paraffin-embedded specimens from bone and ioint tuberculosis （BJTB）. Methods Ninety three BJTB specimens and 12 specimens of bone neoplasm were collected during October 2014 to April 2015 in Beijing Chest Hospital, Capital Medical University. Comparison of FQ-PCR technique and conventional acid-fast staining technique in the diagnosis of tuberculosis in sensitivity and specificity by chi square test. To ob- serve the correlation between the different sits and two detection positive rate, and the non tuberculosis mycobacterium was check out by PCR reverse dot blot hybridization in 7 specimens which had negative mycobacterium DNA and acid-fast staining positive. Results BJTB tissues in FQ-PCR detection of Mycobacterium tuberculosis were positive in 77 patients, positive rate of 82.8% ; 64 cases were acid fast staining positive, positive rate of 68.8%. The sensi tivity of FQPCR method was significantly higher than that of acid-fast staining （X2= 5. 659,P=0. 021）. In different on set location FQ-PCR sensitivity were higher than that of acid fast staining： in the spine, FQ-PCR technique and acid fast staining positive rate was 84. 0% （42/50） and 64.0% （32/50） （X2 =0. 009,P=0. 609） respectively; in the peripheral BJTB, FQ-PCR and acid fast staining positive rate were 81.4% （35/43） and 74.4% （32/43） （Z2 = 12. 609,P = 0. 002） respectively. A strain of mycobacterium Gordon was checked out by PCR-reverse dot blot hybridization. Conclusion FQ-PCR technique improved the positive detection rate of Mycobacterium tuberculosis significantly compared to acid-fast staining and provide important tips for testing non tuberculosis mycobacterium.The FQPCR technology has good application value in the detection of paraffin-embedded specimens from BJTB.