Rapid diagnosis of pulmonary tuberculosis and detection of drug resistance by combined simultaneous amplification testing and reverse dot blot

Abstract

AimsMaking a correct and rapid diagnosis is essential for managing pulmonary tuberculosis (PTB), particularly multidrug-resistant tuberculosis. We aimed to evaluate the efficacy of the combination of simultaneous amplification testing (SAT) and reverse dot blot (RDB) for the rapid detection of Mycobacterium tuberculosis (MTB) and drug-resistant mutants in respiratory samples.Methods225 suspected PTB and 32 non-TB pulmonary disease samples were collected. All sputum samples were sent for acid-fast bacilli smear, SAT, culture and drug susceptibility testing (DST) by the BACTECTM MGITTM 960 system. 53 PTB samples were tested by both RDB and DNA sequencing to identify drug resistance genes and mutated sites.ResultsThe SAT positive rate (64.9%) was higher than the culture positive rate (55.1%), with a coincidence rate of 83.7%. The sensitivity and specificity of SAT for diagnosing PTB were 66.7% and 100%, respectively, while those for culture were 53.9% and 84.2%, respectively. RDB has high sensitivity and specificity in identifying drug resistance genes and mutated sites. The results of RDB correlated well with those of DST and DNA sequencing, with coincidence rates of 92.5% and 98.1%, respectively.ConclusionsThe combination of SAT and RDB is promising for rapidly detecting PTB and monitoring drug resistance in clinical laboratories.

Application of real-time simultaneous amplification and testing method to accurately and rapidly detect extra-pulmonary tuberculosis

“…Simultaneous Amplification and Testing (SAT) is one of the molecular biological detection methods used by the tuberculosis laboratory in China in recent years. It is mainly used to detect MTB in sputum to assist diagnosis [4][5][6][7]. However, SAT has rarely been used to test EPTB specimens and therefore, its sensitivity and specificity for EPTB diagnosis are still not clear.…”

Comparing Mycobacterium tuberculosis RNA Accuracy in Various Respiratory Specimens for the Rapid Diagnosis of Pulmonary Tuberculosis

“… 13 Although most current molecular assays target Mtb DNA, 14 , 15 research on molecular assays that target Mtb RNA has been limited. 16 Shanghai Rendu Biotechnology Co has developed a TB-RNA detection technology that involves simultaneous amplification and testing for the detection of Mtb RNA by employing isothermal 16S ribosomal RNA (rRNA) amplification. 17 This TB-RNA assay has been used for both PTB and EPTB and has shown a relatively good diagnostic accuracy; 18 , 19 however, the total number of studies remains limited.…”

Application of Real-Time Simultaneous Amplification and Testing Method to Accurately and Rapidly Detect Extra-pulmonary Tuberculosis

“…Simultaneous Amplification and Testing (SAT) is one of the molecular biological detection methods used by the tuberculosis laboratory in China in recent years. It is mainly used to detect Mycobacterium tuberculosis (MTB) in sputum to assist diagnosis (Cui Z et al 2012;Zhang Q et al 2017;Yan L et al 2017;Chen Y et al 2018). However, SAT has rarely been used to test EPTB specimens and therefore, its sensitivity and specificity for EPTB diagnosis are still not clear.…”