实时荧光核酸恒温扩增检测技术在结核病诊断中的临床应用

doi:10.3969/j.issn.1000-6621.2014.06.010

摘要/Abstract

摘要： 目的评估实时荧光核酸恒温扩增检测技术（simultaneous amplification and testing，SAT）在结核病患者的痰液、体液及病灶组织等标本中检测结核分枝杆菌的价值。方法收集在广州市胸科医院就诊的疑似结核病患者的痰液734份、体液标本94份（包括68份胸腔积液、21份脑脊液、5份关节积液，以下统称“体液”）及病灶组织标本76份，共计904份标本。同时采用SAT法、改良罗氏培养法进行检测，培养阳性者进行基因芯片菌种鉴定。SAT法与改良罗氏培养法结果不相符的标本，用结核分枝杆菌聚合酶链（polymerase chain reaction technology for Mycobacterium tuberculosis，PCR-TB）荧光诊断试剂盒进行检测。采用χ²检验比较SAT法与改良罗氏培养法对结核病患者的阳性检出率。结果以改良罗氏培养结果为金标准，则SAT法在痰液、体液、病灶组织标本的敏感度、特异度、符合率分别为90.20%（230/255）、92.48%（443/479）、91.69%（673/734）；94.74%（18/19）、81.33%（61/75）、84.04%（79/94）；100.00%（14/14）、77.42%（48/62）、81.58%（62/76）。以扩大金标准（培养+PCR法）比对，则SAT法在痰液、体液、病灶组织标本的敏感度、特异度、符合率分别为96.30%（260/270）、99.35%（461/464）、98.23%（721/734）；96.97%（32/33）、100.00%（61/61）、98.94%（93/94）；100.00%（27/27）、97.96%（48/49）、98.68%（75/76）。改良罗氏培养法和SAT法在痰液、体液、病灶组织标本的阳性检出率分别为34.74%(255/734)和36.24%(266/734)、20.21%(19/94)和34.04%(32/94)、18.42%(14/76)和36.84%(28/76)；痰标本两者的阳性率比较，差异没有统计学意义(χ²=0.36，P＞0.05)。体液与病灶组织标本中,SAT法较改良罗氏培养法的阳性率高出13.83%和18.42%,差异有统计学意义（χ²值分别为4.55、6.44，P值均＜0.05）。结论 SAT法检测痰液、体液及病灶组织标本中的结核分枝杆菌,具有快速、敏感度和特异度较高的优点，可提高结核分枝杆菌的检出率。

关键词: 结核/诊断, 核酸扩增技术, 敏感性与特异性

Abstract: Objective To assess the clinical value of the isothermal RNA amplification assay (SAT) for detection of Mycobacterium tuberculosis in sputum, body fluid and nidus samples． Methods Seven hundred and thirty-four sputum samples, 94 samples of body fluid (including 68 thoracic effusion, 21 cerebrospinal fluid and 5 joint effusion) and 76 nidus samples collected from suspected tuberculosis patients were tested by both SAT and L-J culture. The positive isolates were identified by Gene-chip method. The samples with different results between SAT and L-J culture were tested by Mycobacterium tuberculosis PCR (PCR-TB) fluorescence diagnosis kits．The detection rates of SAT and L-J culture were analyzed by Chi-square test. Results With the result of L-J culture as the re-ference, the sensitivity, the specificity and the coincidence rate of the sputum samples were 90.20% (30/255), 92.48% (443/479) and 91.69% (673/734), those of the fluid samples were 94.74% (18/19), 81.33% (61/75) and 84.04% (79/94), and those of the nidus samples were 100.00% (14/14), 77.42% (48/62) and 81.58% (62/76). With the result of PCR-TB as the reference, the sensitivity, the specificity and the accordance rate of the sputum samples were 96.30% (260/270), 99.35% (461/464) and 98.23% (721/734), those of the fluid samples were 96.97% (32/33), 100.00% (61/61) and 98.94% (93/94), and those of the nidus samples were 100.00% (27/27), 97.96% (48/49) and 98.68% (75/76).The detection rates the three different kinds of specimens by L-J culture and SAT were 34.74% (255/734) vs 36.24% (266/734), 20.21% (19/94) vs 34.04% (32/94), 18.42% (14/76) vs 36.84%(28/76). There was no difference in sputum samples between SAT and L-J culture (χ²=0.36, P＞0.05). However, there were significant differences in body fluids and nidus samples between SAT and L-J culture (χ²=4.55 and 6.44, all P<0.05). Conclusion SAT is a rapid, sensitive and specific method for detection of Mycobacterium tuberculosis in kinds of clinical samples.

Key words: Tuberculosis/diagnosis, Nucleic acid amplification techniques, Sensitivity and specificity

蔡杏珊，马品云，张院良，谭耀驹. 实时荧光核酸恒温扩增检测技术在结核病诊断中的临床应用[J]. 中国防痨杂志, 2014, 36(6): 458-461. doi: 10.3969/j.issn.1000-6621.2014.06.010

CAI Xing-shan, MA Pin-yun, ZHANG Yuan-liang, TAN Yao-ju. Evaluation of the simultaneous amplification and testing for diagnosis of Mycobacterium tuberculosis[J]. Chinese Journal of Antituberculosis, 2014, 36(6): 458-461. doi: 10.3969/j.issn.1000-6621.2014.06.010

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