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
Abstract: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 pulmon… Show more
“…Recently, several new agents were evaluated for digestion and decontamination. Cetylpyridinium chloride (CPC) and sodium chloride (NaCl) methods were confirmed to have a higher detection rate of TB than that of the NALC-NaOH method [ 9 ]. The use of sulfuric acid as the decontamination agent, instead of NaOH, has equal performance in TB detection with the NALC-NaOH method [ 6 ].…”
Section: Discussionmentioning
confidence: 99%
“…During the past decades, various digestion and decontamination methods (DDMs) have been evaluated for sputum culture. However, not much progress has been made [ 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 ].…”
Mycobacterial culture remains the gold standard for the diagnosis of active tuberculosis. However, an appropriate digestion and decontamination method is essential for the effective recovery of tubercle bacilli in culture. The study was designed to compare the efficacy of sputum treated with power ultrasound (PU) and routine NALC-NaOH methods for mycobacterial culture from clinically suspected cases of pulmonary tuberculosis. To evaluate the PU and routine NALC-NaOH methods, sputum specimens (n = 597) were studied (culturing on MGIT 960), and the performances were compared. Of the 597 samples, 89 (14.91%) sputum samples treated with the NaOH-NALC method were mycobacterial culture positive, including Mycobacterium tuberculosis (M.TB; n = 77, 12.90%) and nontuberculous mycobacteria (NTM; n = 12, 2.01%). One hundred and ten (18.43%) sputum samples treated with the PU method were culture positive, including M.TB (n = 87, 14.57%) and NTM (n = 23, 3.85%). The PU method detected 10 additional cases of M.TB and 11 additional cases of NTM when compared to the NALC-NaOH method. Statistical analysis showed that a significant difference was found in the culture-positive ratio of M.TB and NTM between the two method groups (p < 0.05). Compared with that of the NALC-NaOH method (8.04%), sputum treated with PU method (4.86%) had a significantly lower contamination rate (p < 0.05). In conclusion, our data indicate that, compared with the NALC-NaOH method, the PU method is a rapid and effective approach for mycobacterial culture when detecting active TB. However, its accurate mechanism has not been well addressed, and further investigation is still required.
“…Recently, several new agents were evaluated for digestion and decontamination. Cetylpyridinium chloride (CPC) and sodium chloride (NaCl) methods were confirmed to have a higher detection rate of TB than that of the NALC-NaOH method [ 9 ]. The use of sulfuric acid as the decontamination agent, instead of NaOH, has equal performance in TB detection with the NALC-NaOH method [ 6 ].…”
Section: Discussionmentioning
confidence: 99%
“…During the past decades, various digestion and decontamination methods (DDMs) have been evaluated for sputum culture. However, not much progress has been made [ 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 ].…”
Mycobacterial culture remains the gold standard for the diagnosis of active tuberculosis. However, an appropriate digestion and decontamination method is essential for the effective recovery of tubercle bacilli in culture. The study was designed to compare the efficacy of sputum treated with power ultrasound (PU) and routine NALC-NaOH methods for mycobacterial culture from clinically suspected cases of pulmonary tuberculosis. To evaluate the PU and routine NALC-NaOH methods, sputum specimens (n = 597) were studied (culturing on MGIT 960), and the performances were compared. Of the 597 samples, 89 (14.91%) sputum samples treated with the NaOH-NALC method were mycobacterial culture positive, including Mycobacterium tuberculosis (M.TB; n = 77, 12.90%) and nontuberculous mycobacteria (NTM; n = 12, 2.01%). One hundred and ten (18.43%) sputum samples treated with the PU method were culture positive, including M.TB (n = 87, 14.57%) and NTM (n = 23, 3.85%). The PU method detected 10 additional cases of M.TB and 11 additional cases of NTM when compared to the NALC-NaOH method. Statistical analysis showed that a significant difference was found in the culture-positive ratio of M.TB and NTM between the two method groups (p < 0.05). Compared with that of the NALC-NaOH method (8.04%), sputum treated with PU method (4.86%) had a significantly lower contamination rate (p < 0.05). In conclusion, our data indicate that, compared with the NALC-NaOH method, the PU method is a rapid and effective approach for mycobacterial culture when detecting active TB. However, its accurate mechanism has not been well addressed, and further investigation is still required.
“…Recently, various DDMs have been investigated and most of these DDMs use a chemical agent for digestion and the other as a disinfectant [2,[4][5][6][7][8][9][10]. The most commonly used NALC-NaOH digestion and decontamination technique uses 0.5% NALC as a mucolytic agent and NaOH (final concentration of 1%) as a decontaminating agent [14].…”
Section: Discussionmentioning
confidence: 99%
“…Currently, N-Acetyl L Cysteine (NALC)-sodium hydroxide (NaOH) is the extensively used pretreatment technique to isolate MTB from clinical specimens. However, the literature indicates that pre-treatment of clinical samples with NaOH may kill a significant number of tubercle bacilli along with other contaminating organisms [2,4,5].…”
Section: Introductionmentioning
confidence: 99%
“…Several pre-treatment methods have been investigated to isolate mycobacteria from sputum samples [2,[4][5][6][7][8][9][10][11]. However, the complexity and low diagnostic accuracy of these methods limited their use in mycobacterial laboratories [6,9].…”
The present study evaluated the performance of newly developed pancreatin-cetylpyridinium chloride (pancreatin-CPC) digestion and decontamination method (DDM) with N-acetyl L-Cysteine-sodium hydroxide (NALC-NaOH) DDM for isolation of Mycobacteria from clinically suspected pulmonary tuberculosis (PTB) patients. For the study, sputum samples (n = 613) obtained from clinically suspected PTB cases were subjected to direct microscopy, pretreatment with NALC-NaOH DDM (reference method), and pancreatin-CPC DDM followed by culture, and the data were analyzed. The direct microscopy illustrated diagnostic accuracies of 60.4% (sensitivity), 99.77% (specificity), 98.9% (positive predictive value) and 88.3% (negative predictive value), respectively (against culture) for the detection of Mycobacterial species. The pancreatin-CPC DDM showed competitive diagnostic accuracies (against NALC-NaOH DDM) of 99.32% (sensitivity), 94.07% (specificity), 85.05% (positive predictive value), and 99.76% (negative predictive value), respectively, for the isolation of Mycobacterial species. In conclusion, pancreatin-CPC DMM was a highly sensitive, technically simple, and cost-effective method, suggesting its competence to substitute the currently used NALC-NaOH DDM.
Antibiotic resistance continues to be a most serious threat to public health. This situation demands that the scientific community increase their efforts for the discovery of alternative strategies to circumvent the problems associated with conventional small molecule therapeutics. The Global Antimicrobial Resistance and Use Surveillance System (GLASS) Report (published in June 2021) discloses the rapidly increasing number of bacterial infections that are mainly caused by antimicrobial-resistant bacteria. These concerns have initiated various government agencies and other organizations to educate the public regarding the appropriate use of antibiotics. This review discusses a brief highlight on the timeline of antimicrobial drug discovery with a special emphasis on the historical development of antimicrobial resistance. In addition, new antimicrobial targets and approaches, recent developments in drug screening, design, and delivery were covered. This review also discusses the emergence and roles of various antibiotic adjuvants and combination therapies while shedding light on current challenges and future perspectives. Overall, the emergence of resistant microbial strains has challenged drug discovery but their efforts to develop alternative technologies such as nanomaterials seem to be promising for the future.
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