Comparison of N-acetyl-L-cysteine-sodium hydroxide and Modified Petroff’s Decontamination Methods for Mycobacterium tuberculosis Culture

Ariyani Kiranasari, M. Rayhan
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Abstract

Indonesia is one of 22 countries with a high incidence of TB in the world, particularly related to TB-HIV and MDR-TB cases. Contamination of  normal flora from nasopharyngeal tract is the main problem  to isolate Mycobacterium tuberculosis (MTB) from sputum. It is needed a safe solution to decontaminate without killing  MTB bacilli. N-acetyl-L-cysteine-sodium hydroxide (NALC-NaOH) is compared with  modified Petroff’s  which is widely used in laboratories, to get better outcome. Of the 110 sputum samples were collected from suspected cases of Pulmonary TB. Decontamination using 2% NALC-NaOH and Modifed Petroff  (4% NaOH) was performed before AFB smear and culture on LJ medium. Each group was assessed for contamination and culture positive rate. The positive culture was validated using chromatography test for detected antigen MPT-64 and PNB. Result of this study showed that NALC-NaOH and  modified Petroff’s  methods did not significantly affect positivity rate of  Acid-Fast Bacillus (AFB) smear, 71% and 66% respectively. Contamination on culture was significantly (p=0,034) higher in samples treated with NALC-NaOH (21%) compared to Modified Petroff methods(13%). Proportion of positive culture on samples treated with NALC-NaOH was lower than Modified Petroff, 65% and 70% respectively with p value=1. Conclusion of this study showed that sputum  decontamination using Modified Petroff methods is  still more effective than  NALC-NaOH  to increase  positivity rate of MTB culture.  However, the two methods were not significantly different to get positivity result on the microscopic examination  of AFB.
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n -乙酰- l-半胱氨酸-氢氧化钠与改良Petroff去污方法对结核分枝杆菌培养效果的比较
印度尼西亚是世界上22个结核病高发国家之一,特别是与结核病-艾滋病毒和耐多药结核病病例有关。从痰液中分离结核分枝杆菌(MTB)的主要问题是鼻咽部正常菌群污染。需要一种不杀死结核分枝杆菌的安全的去污溶液。将n -乙酰- l-半胱氨酸-氢氧化钠(NALC-NaOH)与实验室中广泛使用的改性Petroff进行比较,得到更好的结果。其中110份痰样本采集自疑似肺结核病例。使用2% NALC-NaOH和改性Petroff (4% NaOH)进行净化,然后在LJ培养基上进行AFB涂片和培养。评估各组污染及培养阳性率。采用层析法检测抗原MPT-64和PNB,验证阳性培养。本研究结果表明,NALC-NaOH和改进的Petroff方法对抗酸芽孢杆菌(AFB)涂片的阳性率无显著影响,分别为71%和66%。与改良的Petroff方法(13%)相比,NALC-NaOH处理的样品(21%)对培养物的污染显著(p= 0.034)更高。NALC-NaOH处理样品的阳性培养比例低于Modified Petroff,分别为65%和70%,p值=1。本研究结论表明,改良Petroff法对痰液去污仍比NALC-NaOH法对提高MTB培养阳性率更有效。两种方法镜检AFB阳性结果差异无统计学意义。
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