Ustinova Vera V, Smirnova Tatiana G, Sochivko Dmitry G, Varlamov Dmitry A, Larionova Elena E, Andreevskaya Sofya N, Andrievskaya Irina Yu, Kiseleva Ekaterina A, Chernousova Larisa N, Ergeshov Atadzhan
Microbiology Department, Central Tuberculosis Research Institute RAMS, Moscow, Russia.
Microbiology Department, Central Tuberculosis Research Institute RAMS, Moscow, Russia.
Tuberculosis (Edinb). 2019 Jan;114:17-23. doi: 10.1016/j.tube.2018.10.004. Epub 2018 Oct 9.
The purpose of the present study was to create a real-time PCR test system allowing simultaneous detection of nontuberculous mycobacteria (NTM) and Mycobacterium tuberculosis complex (MTBC) both in culture and sputum. NTM cultures (18 strains, 18 species), MTBC cultures (16 strains, 2 species) and non-mycobacterial microorganisms from the collection of the Central Research TB Institute (CTRI) were used for the preliminary evaluation of the test system. 301 NTM cultures from patients with mycobacteriosis were used to assess the sensitivity of the developed test system. Clinical respiratory samples (sputum) from 104 patients with mycobacteriosis, 3627 patients with tuberculosis and 118 patients with other lung diseases were used for diagnostic sensitivity and specificity testing. The specificity and sensitivity of the assay for MTBC was found to be 100% both in culture and sputum samples; for NTM, the specificity was 100% in culture and sputum, the sensitivity reached 100% in culture and 73.1% in sputum samples. Positive predictive value (PPV) and negative predictive value (NPV) of the assay for culture were both 100%, for clinical material 100% and 80.8%, respectively. The limit of detection at the probability of detection 95% (LoD95%) was estimated to be 16 cfu/ml for M. tuberculosis H37RV and 1200 cfu/ml for M. avium.
本研究的目的是创建一种实时PCR检测系统，能够同时在培养物和痰液中检测非结核分枝杆菌（NTM）和结核分枝杆菌复合群（MTBC）。使用中央结核病研究所（CTRI）收集的NTM培养物（18株，18种）、MTBC培养物（16株，2种）和非分枝杆菌微生物对该检测系统进行初步评估。使用来自分枝杆菌病患者的301份NTM培养物评估所开发检测系统的灵敏度。来自104例分枝杆菌病患者、3627例结核病患者和118例其他肺部疾病患者的临床呼吸道样本（痰液）用于诊断灵敏度和特异性检测。结果发现，该检测方法对MTBC的特异性和灵敏度在培养物和痰液样本中均为100%；对于NTM，培养物和痰液中的特异性均为100%，培养物中的灵敏度达到100%，痰液样本中的灵敏度为73.1%。该检测方法对培养物的阳性预测值（PPV）和阴性预测值（NPV）均为100%，对临床样本的阳性预测值和阴性预测值分别为100%和80.8%。估计结核分枝杆菌H37RV在检测概率为95%时的检测限（LoD95%）为16 cfu/ml，鸟分枝杆菌为1200 cfu/ml。
