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Journal of Clinical Microbiology, July 1998, p. 1996-2003, Vol. 36, No. 7
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Novel Method for Processing Respiratory Specimens for Detection of Mycobacteria by Using C18-Carboxypropylbetaine: Blinded Study

Charles G. Thornton,1,* Kerry M. MacLellan,1 Thomas L. Brink Jr.,1 Denise E. Lockwood,2 Mark Romagnoli,3 June Turner,4 William G. Merz,3 Richard S. Schwalbe,5 Marcia Moody,4 Yvonne Lue,6 and Selvin Passen1

Department of Molecular Biology and Genetics1 and Department of Microbiology,2 Quest Diagnostics---Baltimore, Baltimore, Maryland 21227; Department of Pathology, Johns Hopkins Medical Institutes, Baltimore, Maryland 212873; District of Columbia Department of Human Services Bureau of Laboratories, Washington, D.C. 200014; Department of Pathology, University of Maryland at Baltimore, Baltimore, Maryland 212015; and Quest Diagnostics---Teterboro, Teterboro, New Jersey 076086

Received 26 September 1997/Returned for modification 11 November 1997/Accepted 7 January 1998

A novel method for processing respiratory specimens to improve culture and acid-fast staining of mycobacteria is introduced. This new method utilized N,N-dimethyl-N-(n-octadecyl)-N-(3-carboxypropyl)ammonium inner salt (Chemical Abstract Service no. 78195-27-4), also known as C18-carboxypropylbetaine (CB-18). In a blinded, five-center study, CB-18-based processing was compared to the standard method combining NALC and NaOH (NALC/NaOH). A total of 573 respiratory specimens were tested. Individual specimens were split approximately equally; the host institutions processed half of each specimen by the NALC/NaOH method, while the other half was processed with CB-18 at Quest Diagnostics---Baltimore. A total of 106 specimens were culture positive for acid-fast bacilli (AFB). Replacement of the primary decontamination agent with CB-18 caused changes in all diagnostic parameters. Aggregate culture sensitivity improved by approximately 43% (P < 0.01), and smear sensitivity improved by approximately 58% (P < 0.01). The sensitivity of smear relative to that of M. tuberculosis isolates exceeded 93% (P < 0.01) when specimens were processed with CB-18. The average times to a positive result were reduced by 7.3 days in liquid culture (P < 0.01) and 5.3 days on solid media (P < 0.05); however, the CB-18 method had a 20.8% contamination rate in liquid culture versus a rate of approximately 7.5% with NALC/NaOH processing. There were also unusual reductions in liquid culture sensitivity and smear specificity among CB-18-processed specimens. The characteristics of the latter parameters suggested that refinement of the CB-18 processing method should allow further improvements in culture sensitivity. This study showed that the CB-18 method has the potential to improve both smear and culture detection for these important human pathogens.

* Corresponding author. Mailing address: Quest Diagnostics, Inc., Department of Molecular Biology and Genetics, 1901 Sulphur Spring Rd., Baltimore, MD 21227. Phone: (410) 536-1524. Fax: (410) 536-1633. E-mail: thornton{at}msmail.mml.com.

Journal of Clinical Microbiology, July 1998, p. 1996-2003, Vol. 36, No. 7
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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