河北医科大学学报 ›› 2022, Vol. 43 ›› Issue (1): 39-43.doi: 10.3969/j.issn.1007-3205.2022.01.009

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实时荧光定量聚合酶链反应技术在肾结核病理诊断中的应用价值

  

  1. 河北省胸科医院泌尿外科，河北 石家庄 050000
  • 出版日期:2022-01-25 发布日期:2022-01-24
  • 作者简介:范正超(1984-)，男，河北清河人，河北省胸科医院主治医师，医学博士研究生，从事泌尿生殖系统结核的微创腔镜治疗研究。
  • 基金资助:
    河北省医学科学研究重点课题计划（20170412）

Application value of real-time fluorescent quantitative polymerase chain reaction technique in pathological diagnosis of renal tuberculosis

  1. Department of Urology, Hebei Chest Hospital, Shijiazhuang 050000, China
  • Online:2022-01-25 Published:2022-01-24

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摘要： 目的  探讨实时荧光定量聚合酶链反应（polymerase chain reaction，PCR）技术在肾结核病理诊断中的应用价值。
方法  回顾性收集我院泌尿外科通过后腹腔镜技术或开放技术行肾切除术后石蜡组织标本55例患者的临床资料，以术前尿结核分枝杆菌培养结果为金标准，并结合术后病理诊断结果，将研究对象分为结核肾病组25例（阳性组），非结核肾病组30例（阴性组）。采用Taqman探针法实时荧光定量PCR技术和抗酸染色技术检测每个石蜡组织标本，对比分析两种检测技术在肾结核病理诊断中的诊断效能，绘制受试者工作特征（receiver operating characteristic，ROC）曲线评估两种方法在肾结核病理诊断中的价值。
结果  实时荧光定量PCR技术检测的敏感度与抗酸染色技术检测的敏感度分别为80.0%（20/25）和48.0(12/25)，前者相比后者敏感度提高了32.0%，差异有统计学意义（χ2=5.556，P=0.018）。荧光定量PCR检测的特异度、阳性预测值、阴性预测值均高于抗酸染色检测的结果，但两者比较差异均无统计学意义（P＞0.05）。荧光定量PCR检测的准确度高于抗酸染色检测的准确度（89.1% vs. 72.7%），两者比较差异有统计学意义（χ2=4.767，P=0.029）。两种技术检测肾结核组织标本病原体结果与尿结核分枝杆菌培养结果的一致性Kappa值分别为0.777和0.429。ROC曲线分析显示，实时荧光定量PCR技术和抗酸染色技术的ROC曲线下面积（area under curve,AUC）分别为0.883和0.707,前者高于后者0.177,差异有统计学意义（Z=3.502，P＜0.05）。
结论  实时荧光定量PCR技术检测方法简单易行，与抗酸染色检测技术相比可以提高肾结核组织病原体检测的敏感度和准确度，在肾结核的病理诊断中具有良好的应用价值。


关键词: 结核, 肾, 实时聚合酶链反应, 组织细胞学制备技术

Abstract: Objective  To investigate the application value of real-time fluorescent quantitative polymerase chain reaction(FQ-PCR) technique in pathological diagnosis of renal tuberculosis. 
Methods  We retrospectively analyzed 55 patients with paraffin-embedded specimens after retroperitoneoscopic or open nephrectomy in Department of Urology of our hospital. Taking the results of preoperative urine Mycobacterium tuberculosis culture results as the gold standard, combined with the results of postoperative pathological diagnosis, the subjects were divided into tuberculos nephropathy group(positive group, n=25) and non-tuberculous nephropathy group(negative group, n=30). Each paraffin-embedded tissue specimen was detected using TaqMan-probe real-time FQ-PCR technique and and acid-fast staining technique, and the diagnostic efficiency of the two detection techniques in the pathological diagnosis of renal tuberculosis was compared and analyzed. A recevier operating characteristic(ROC) curve was drawn to evaluate the value of the two methods in the pathological diagnosis of renal tuberculosis. 
Results  The sensitivity of real-time FQ-PCR and acid-fast staining was 80.0%(20/25) and 48.0%(12/25) respectively; the sensitivity of the former was higher by 32.0% than that of acid-fast staining,with significant difference(χ2=5.556， P=0.018). The specificity，the positive predictive value and the negative predictive value of real-time FQ-PCR were higher than that of acid-fast staining, but there was no significant difference between them（P＞0.05）. The accuracy of real-time FQ-PCR was higher than that of acid-fast staining（89.1% vs. 72.7%）, and there was significant difference between them（χ2=4.767， P=0.029）. The Kappa values of the two techniques for detecting pathogens in renal tuberculosis tissue specimens which were consistent with results of urine Mycobacterium tuberculosis culture were 0.777 and 0.429, respectively. ROC curve analysis showed that area under ROC curve(AUC)of real-time FQ-PCR technique and acid-fast staining technique was 0.883 and 0.707 respectively; the former was higher than the latter by 0.177, and the difference was statistically significant(Z=3.502, P＜0.05). 
Conclusion  Real-time FQ-PCR technique is simple and has higher sensitivity and accuracy in the detection of pathogens in renal tuberculosis tissue, as compared with acid-fast staining technique. Real-time FQ-PCR technique shows good application value in pathological diagnosis of renal tuberculosis.


Key words: tuberculosis, renal, real-time Polymerase chain reaction, histocytological preparation techniques

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