Simultaneous amplification and testing method forMycobacterium tuberculosisrRNA to differentiate sputum-negative tuberculosis from sarcoidosis

Abstract

We use the simultaneous application and testing method to detect Mycobacterium tuberculosis rRNA (SAT-TB) with the endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) biopsy specimens to differentiate sputum-negative tuberculosis from sarcoidosis. In the first part, we validated the SAT-TB on the bronchial or EBUS-TBNA biopsy specimens from sputum smear-positive pulmonary tuberculosis. In the second part, all EBUS-TBNA specimens for sputum smear-negative intrathoracic tuberculous lymphadenopathies or sarcoidosis were tested with the SAT-TB, acid-fast bacilli smear, and culture. In the 16 sputum-positive tuberculosis cases, 5 showed negative SAT (2 nontuberculous mycobacteria and 3 had anti-tuberculosis therapies previously); the remaining 11 were positive. Of the 41 sputum-negative tuberculosis cases in the second part, five other diseases were negative. In the remaining 36 cases, 27 sarcoidosis cases were negative; 7 in 9 with sputum-negative tuberculosis were positive (77.78%). In these 36 patients, the sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of the SAT method were 77.78, 100, 100, 93.10, and 94.44%, respectively. The SAT distinguished sputum-negative tuberculosis from sarcoidosis significantly ( P < 0.0001) and identified cases with active M. tuberculosis as accurately as the conventional methods (κ = 0.912, P < 0.0001). We conclude that the SAT-TB may be an effective method for using biopsy specimens to differentiate sputum-negative tuberculosis from sarcoidosis.

Identification of Biomarkers for Sarcoidosis and Tuberculosis of the Lung Using Systematic and Integrated Analysis

“…Pulmonary tuberculosis (PTB) is an infectious disease caused by Mtb that mainly affects the lungs and lymph nodes. SARC and sputum-negative PTB share similar clinical, radiological, and histopathological characteristics, thereby making the differential diagnosis of sputum-negative TB and SARC challenging [ 5 ]. The blood profiles of patients with PTB and SARC predominantly have similar indications of inflammation [ 6 ].…”

Rapid discrimination between tuberculosis and sarcoidosis using next-generation sequencing

“…This may be attributed to sample contamination. A previous meta-analysis indicated that 26% of specimens obtained from SA patients contained MTB nucleic acid contamination (Gupta et al, 2007;Li et al, 2019).…”

“…Studies have shown that in uncultured samples, the concentration of MTB DNA is lower than that of the host DNA, which yields very low coverage and depth of NGS (Iketleng et al, 2018). In addition, the low number of reads may also be due to the contaminated SA samples (Li et al, 2019). We further attempted to carefully remove false-positive results.…”

“…For example, PCR amplification of IS6110 and MPB64 has shown a sensitivity of 78% and a specificity of 100% for MTB identification (Agrawal et al, 2016). The detection of a 170-bp MTB 16S rRNA fragment could even allow the differential diagnosis of sputum-negative TB from SA (Li et al, 2019). Unfortunately, up to 26% of SA specimens contain MTB nucleic acids (Gupta et al, 2007), which poses great challenges in the discrimination of these two diseases.…”

Characteristics and management of ocular sarcoidosis

“…Meanwhile, it is believed that the allergic condition of the patients to certain pathogens plays an important role in granulomatous disease, including sarcoidosis. Various pathogens, such as mycobacteria [15][16][17], mumps virus [18], and human herpes viruses [19] have been listed as candidates for sarcoidosis. While mycobacteria are regarded as an important candidate causative pathogen of sarcoidosis, especially in Western countries, Propionibacterium acnes (P. acnes), currently reclassified as Cutibacterium acnes, has been investigated as the causative agent of sarcoidosis in Japanese groups.…”