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Journal of Clinical Microbiology, June 2005, p. 2703-2708, Vol. 43, No. 6
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.6.2703-2708.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Utility of Universal Sample Processing Methodology, Combining Smear Microscopy, Culture, and PCR, for Diagnosis of Pulmonary Tuberculosis

Soumitesh Chakravorty,1,{dagger} Mridu Dudeja,1,{ddagger} M. Hanif,2 and Jaya Sivaswami Tyagi1*

Department of Biotechnology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029,1 New Delhi Tuberculosis Center, New Delhi 110002, India2

Received 12 February 2004/ Returned for modification 26 July 2004/ Accepted 6 September 2004

The universal sample processing (USP) multipurpose methodology was developed for the diagnosis of tuberculosis (TB) and other mycobacterial diseases by using smear microscopy, culture, and PCR (S. Chakravorty and J. S. Tyagi, J. Clin. Microbiol. 43:2697-2702, 2005). Its performance was evaluated in a blinded study of 571 sputa and compared with that of the direct and N-acetyl L-cysteine (NALC)-NaOH methods of smear microscopy and culture. With culture used as the gold standard, USP smear microscopy demonstrated a sensitivity and specificity of 98.2% and 91.4%, respectively, compared to 68.6% and 92.6%, respectively, for the direct method. For a subset of 325 specimens, the USP method recorded a 97.1% sensitivity and 83.2% specificity compared to the NALC-NaOH method, which had a sensitivity and specificity of 80.0% and 89.7%, respectively, with culture used as the gold standard. Thus, the USP method exhibited a highly significant enhancement in sensitivity (P < 0.0001) compared to the direct and NALC-NaOH methods of smear microscopy. The USP culture sensitivity was 50.1% and was not significantly different from that of conventional methods (53.6%). The sensitivity and specificity of IS6110 PCR were 99.1% and 71.2%, respectively, with culture used as the gold standard, and increased to 99.7% and 78.8%, respectively, when compared with USP smear microscopy. Thus, the USP methodology was highly efficacious in diagnosing TB by smear microscopy, culture, and PCR in a clinical setting.

* Corresponding author. Mailing address: Department of Biotechnology, All India Institute of Medical Sciences, New Delhi 110029, India. Phone: 91-11-26588491. Fax: 91-11-26588663. E-mail: jstyagi{at}aiims.ac.in.

{dagger} Present address: Center for Emerging Pathogens, Department of Medicine, New Jersey Medical School (UMDNJ), Newark, NJ 07103.

{ddagger} Present address: Sunderlal Charitable Trust Hospital, Delhi, India.

Journal of Clinical Microbiology, June 2005, p. 2703-2708, Vol. 43, No. 6
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.6.2703-2708.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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