Document Detail

Simplified acetylcysteine-alkali digestion-decontamination procedure for isolation of mycobacteria from clinical specimens.
MedLine Citation:
PMID:  3305557     Owner:  NLM     Status:  MEDLINE    
During our attempts to reevaluate and improve mycobacteriology laboratory procedures, we found that isolation of mycobacteria from undecontaminated specimens through the use of selective media was unrewarding and that specimen decontamination was an essential step. A reevaluation of the N-acetyl-L-cysteine-sodium hydroxide (NALC-NaOH) decontamination procedure (G. P. Kubica, W. E. Dye, M. L. Cohn, and G. Middlebrook, Am. Rev. Respir. Dis. 87:775-779, 1963) indicated that this method could be simplified and shortened without affecting the isolation rate or growth and without increasing the contamination rate. The modified NALC-NaOH procedure effectively combines the decontamination step with the concentration step: specimens are mixed with the NALC-NaOH solution on a Vortex mixer and, without any waiting or addition of buffer or water to the digested specimens, centrifuged at 3,000 X g for 15 min, and the sediment is suspended in phosphate buffer (pH 5.3). The modified method is simpler, faster, and safer than the original procedure, and the reduced manipulation is also likely to minimize the chance for cross-contamination in sequential specimen processing. The total alkali exposure time in the modified NALC-NaOH method should be kept to a minimum; if necessary or feasible, the concentration of NaOH in the digestion solution may be reduced from the usual strength of 2% to about 1.5% to compensate for the longer alkali exposure time in the modified method to maintain the isolation rate or to improve it.
S Ratnam; F A Stead; M Howes
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of clinical microbiology     Volume:  25     ISSN:  0095-1137     ISO Abbreviation:  J. Clin. Microbiol.     Publication Date:  1987 Aug 
Date Detail:
Created Date:  1987-10-21     Completed Date:  1987-10-21     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505564     Medline TA:  J Clin Microbiol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1428-32     Citation Subset:  IM    
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MeSH Terms
Bacteriological Techniques*
Culture Media
Hydrogen-Ion Concentration
Mycobacterium / isolation & purification*
Sodium Hydroxide*
Sputum / microbiology
Urine / microbiology
Reg. No./Substance:
0/Culture Media; 1310-73-2/Sodium Hydroxide; 616-91-1/Acetylcysteine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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