Use of Pleural Fluid Digital PCR Analysis to Improve the Diagnosis of Pleural Tuberculosis

Abstract

Pleural TB is one of the most frequent causes of pleural effusion, especially in areas with high burden of TB. Due to the paucibacillary nature of the disease, the diagnostic sensitivities of all available bacteriological and molecular tests remain poor.

The incremental value of Mycobacterium tuberculosis trace nucleic acid detection in CT-guided percutaneous biopsy needle rinse solutions for the diagnosis of tuberculosis

“…Conserved DNA sequences in M.tb complex, insertion sequence IS 6110 and IS 1081 , were used as detection targets in this study. The primers for amplification, oligonucleotide probes and dPCR assay procedures have been previously optimized and described in Li et al ( 2022 ). The 20-μL reaction mixtures (10 μL ddPCR TM supermix for probes, 0.9 μM each primer, 0.2 μM each probe, 0.3 U uracil-N-glycosylase, DNA samples without dilution) and 70-μL droplet generation oil were added in cartridges and droplets were generated using QX200 Droplet Generator (Bio-Rad).…”

“…Compared to qPCR, dPCR offers advantages such as quantification without the need for a standard curve, improved precision, and enhanced tolerance to PCR inhibitors (Kuypers and Jerome, 2017 ). It has demonstrated good efficacy in improving the diagnosis of paucibacillary TB, including tuberculous meningitis and pleurisy (Li et al, 2020 , 2022 ). The aim of this study was to assess the value of computed tomography (CT) - guided percutaneous BNRS-dPCR analysis in the diagnosis of TB.…”

Developing a method to detect lipoarabinomannan in pleural fluid and assessing its diagnostic efficacy for tuberculous pleural effusion

“…Patients were divided into three groups according to diagnostic criteria [ 14 , 15 ]：(1) Definite TPE: Mycobacterium tuberculosis (Mtb) is isolated from PF, sputum, or pleural tissue through culture, microscopic examination, or Gene-Xpert, or pleural biopsy shows caseous granulomas. (2) Probable TPE: Pathogens are not confirmed in PF, but all patients are empirically treated for tuberculosis based on clinical suspicion (such as typical clinical symptoms, significant radiographic/pleural effusion, elevated PF adenosine deaminase (ADA), and positive immunological results (interferon-gamma release assays or tuberculin skin test)).…”

Tuberculous pleuritis: clinical presentations and diagnostic challenges

“…These contemporary sequencing techniques can directly confirm the presence of TB bacilli in the pleural space by identifying their specific gene fragments. Again, the lack of a gold standard means that the performances of these technologies are lower if a composite endpoint, instead of positive culture results, was taken as the final verdict [ 56 , 57 , 58 ▪▪ , 59 , 60 ]. There is thus no consensus on how these techniques are best applied clinically.…”