Papain-cetylpyridinium chloride and pepsin-cetylpyridinium chloride; two novel, highly sensitive, concentration, digestion and decontamination techniques for culturing mycobacteria from clinically suspected pulmonary tuberculosis cases

• Published in: PloS one 2020-08, Vol.15 (8), p.e0236700-e0236700
• Main Authors: Shinu, Pottathil, Singh, Varsha A, Nair, Anroop, Venugopala, Katharigatta N, Akrawi, Sabah H
• Format: Article
• Language: English
• Subjects: Antiseptics; Bacilli; Bacterial cultures; Biology and Life Sciences; Cetylpyridinium chloride; Chloride; Chlorides; Decontamination; Diagnosis; Digestion; Enzymes; Funding; Glycerol; Laboratories; Light emitting diodes; Media (culture); Medicine and Health Sciences; Methods; Mycobacterium tuberculosis; Papain; Pepsin; Pharmaceutical sciences; Pharmacy; Pulmonary tuberculosis; Recovery; Research and Analysis Methods; Sensitivity; Sodium; Sodium hydroxide; Software; Sputum; Testing; Tuberculosis
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0236700

Mycobacterial culture remains the gold standard for the diagnosis of tuberculosis. However, an appropriate digestion and decontamination method (DDM) is essential for the effective recovery of tubercle bacilli in culture. Therefore, the current study was designed to compare the performance of papain-cetylpyridinium chloride [papain-CPC] and pepsin-cetylpyridinium chloride [pepsin-CPC] DDMs against N-acetyl L-Cysteine-sodium hydroxide (NALC-NaOH) DDM for recovery of mycobacteria from clinically suspected pulmonary tuberculosis cases. To evaluate papain-CPC, pepsin-CPC and NALC-NaOH DDMs, sputum samples (N = 1381) were cultured on Löwenstein-Jensen medium and the results were compared. The papain-CPC DDM showed sensitivity, specificity, positive predictive value, and negative predictive value of 100%, 93.27%, 71.7%, and 100%, respectively as compared to NALC-NaOH DDM. Similarly, pepsin-CPC DDM demonstrated sensitivity, specificity, positive predictive value and negative predictive value of 98.94%, 94.7%, 76.11%, and 99.81%, respectively. In summary, both papain-CPC and pepsin-CPC DDMs are highly sensitive and specific techniques for recovery of mycobacteria as compared to NALC-NaOH DDM. However, when the overall performances of all DDMs compared, papain-CPC DDM isolated increased number of mycobacterial isolates with comparatively higher numbers of colonies on LJ media than both pepsin-CPC and NALC-NaOH DDMs, indicating its potential to replace the NALC-NaOH DDM for recovery of mycobacteria from sputum samples.