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Journal of Clinical Microbiology, December 1998, p. 3558-3566, Vol. 36, No. 12
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

In Vitro Comparison of NALC-NaOH, Tween 80, and C18-Carboxypropylbetaine for Processing of Specimens for Recovery of Mycobacteria

Charles G. Thornton,* Kerry M. MacLellan, Thomas L. Brink Jr.,dagger and Selvin Passen

Integrated Research Technology, LLC, Baltimore, Maryland 21227

Received 3 August 1998/Returned for modification 3 September 1998/Accepted 22 September 1998

A recent article (C. G. Thornton et al., J. Clin. Microbiol. 36:1996-2003, 1998) reported a new specimen-processing method for improved recovery of mycobacteria. This method used C18-carboxypropylbetaine (CB-18) and increased both smear and culture sensitivity. The companion article (C. G. Thornton et al., J. Clin. Microbiol. 36:2004-2013, 1998) described initial improvements to this method. Additional significant parameters of the CB-18 processing method are identified herein. First, eliminating the incubation step was shown to further improve culture sensitivity. Subsequently, recovery of several mycobacterial isolates by the CB-18 method was compared to a contemporary processing method that combines NALC and NaOH (NALC-NaOH) and a Tween 80-based method. Recovery of the tuberculous isolates following NALC-NaOH processing averaged 20% and ranged from 1.6 to 45%, whereas recovery of the nontuberculous isolates averaged 11% and ranged from 0.1 to 55%. Recovery of the tuberculous and nontuberculous isolates by the Tween 80-based method ranged from 22 to 92% and 27 to 93%, respectively, with averages of 58 and 65%, respectively. Recovery of the tuberculous and nontuberculous mycobacteria following CB-18 processing averaged 86 and 73%, respectively, with ranges from 61 to over 100% and from 43 to over 100%, respectively. Other parameters of the CB-18 method were also examined, including recovery versus CB-18 concentration and the relationship between CB-18 concentration and the tuberculocidal effect. The tuberculocidal effect was time dependent but independent of concentration, whereas recovery was directly proportional to concentration. Increasing the CB-18 concentration to 4 mM provided quantitative recovery on solid medium; however, higher concentrations of CB-18 were not compatible with liquid culture. Examination of the relationship between increasing CB-18 and lecithin concentrations suggested that lecithin could not overcome the deleterious effects of CB-18 in liquid culture at these higher concentrations.

* Corresponding author. Mailing address: Integrated Research Technology, LLC, c/o Quest Diagnostics Incorporated, 1901 Sulphur Spring Rd., Baltimore, MD 21227. Phone: (410) 536-1524. Fax: (410) 536-1633. E-mail: 104217.456{at}compuserve.com.

dagger Present address: Becton Dickinson Advanced Diagnostics, Sparks, MD 21152.

Journal of Clinical Microbiology, December 1998, p. 3558-3566, Vol. 36, No. 12
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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