Rapid Detection of Mycobacterium tuberculosis in Pleural Fluid Using Resuscitation-Promoting Factor-Based Thin Layer Agar Culture Method.

Fengjiao Du,Qi Sun,Zhongquan Liu,Boping Du,Liping Pan,Rongrong Wei,Zihui Li,Hongyan Jia,Aiying Xing,Zongde Zhang

doi:10.3389/fmicb.2022.803521

Abstract

BackgroundPleural tuberculous is difficult to diagnose. Culture is still considered the gold standard, especially in resource-limited settings where quick, cheap, and easy techniques are needed. The aim of the study was to evaluate resuscitation-promoting factors (Rpfs)-based thin layer agar (TLA) culture method for quick detection of Mycobacterium tuberculosis in pleural fluid.MethodsPatients with suspected pleural TB were enrolled prospectively in our hospital, pleural fluid of all patients were collected, stained with Ziehl–Neelsen for acid-fast bacilli (AFB), cultured on Rpfs-TLA, TLA, and Löwenstein–Jensen (LJ) medium, and identified according to recommended procedures.ResultsA total of 137 suspected pleural TB were enrolled and categorized, including 103 pleural TB (49 confirmed and 54 probable pleural TB) and 34 non-TBP patients. The sensitivity of Rpfs-TLA for total pleural TB was 43.7% (34.5∼53.3%), higher than that of TLA 29.1% (21.2∼38.5%) and LJ 26.2% (18.7∼35.5%) (p < 0.01), and all specificity was 100% in the diagnosis of pleural TB. Median time to detection of a positive culture was 11.8 days (95% CI 10.4∼13.4) for Rpfs-TLA, 21.0 days (95% CI 19.1∼22.9) for TLA, and 30.5 days (95% CI 28.5∼32.5) for LJ (p < 0.001).ConclusionRpfs-TLA is an accurate, rapid, cheap, and easy culture method, which makes it promising for use in clinical laboratories.

Highlights

Tuberculosis (TB) is one of the infectious diseases with the highest morbidity and mortality in the world, with an estimated 9.87 million new cases and 1.50 million deaths in 2020
Median time to detection of a positive culture was 11.8 days for resuscitationpromoting factors (Rpfs)-thin layer agar (TLA), 21.0 days for TLA, and 30.5 days for LJ (p < 0.001)
Rpfs-TLA is an accurate, rapid, cheap, and easy culture method, which makes it promising for use in clinical laboratories

Summary

Introduction

Tuberculosis (TB) is one of the infectious diseases with the highest morbidity and mortality in the world, with an estimated 9.87 million new cases and 1.50 million deaths in 2020. Due to the paucibacillary nature of the disease and/or non-replicating dormant bacteria that exists, reducing growth potential of these organisms in pleural effusion (PE) (Mukamolova et al, 2010; Rosser et al, 2017; Shaw et al, 2018; Dusthackeer et al, 2019), traditional microbiological methods are poor and long time consuming, for example, microscopy of pleural fluid and Ziehl–Neelsen staining are rarely positive (0–5%), and culture has more sensitivity of 24–58%, but takes 4– 8 weeks to return a result (Barbas et al, 1991; Gopi et al, 2007; Trajman et al, 2008). The aim of the study was to evaluate resuscitationpromoting factors (Rpfs)-based thin layer agar (TLA) culture method for quick detection of Mycobacterium tuberculosis in pleural fluid

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