Clinical Usefulness of Real-time PCR and Amplicor MTB PCR Assays for Diagnosis of Tuberculosis

Chae Lim Jung; Mi Kyung Kim; Dong Chun Seo; Mi Ae Lee

doi:10.5145/kjcm.2008.11.1.29

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Jung, Kim, Seo, and Lee: Clinical Usefulness of Real-time PCR and Amplicor MTB PCR Assays for Diagnosis of Tuberculosis

Original Article

Korean J Clin Microbiol 2008;11(1):29-33.

Published online: 20 April 2008

DOI: https://doi.org/10.5145/kjcm.2008.11.1.29

Clinical Usefulness of Real-time PCR and Amplicor MTB PCR Assays for Diagnosis of Tuberculosis

Chae Lim Jung, Mi Kyung Kim, Dong Chun Seo, Mi Ae Lee

Department of Laboratory Medicine, School of Medicine, Ewha Womans University Mokdong Hospital, Seoul, Korea

Mi Ae Lee, Department of Laboratory Medicine, Ewha Womans University Mokdong Hospital, 911-1, Mok-dong, Yangcheon-gu, Seoul, 158-056, Korea. (Tel) 82-2-2650-5222, (Fax) 82-2-2650-5222, (E-mail) miae@ewha.ac.kr

Received 1 February 2008 Accepted 4 March 2008

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

PCR assay has provided a mean of more rapid and sensitive detection of Mycobacterium tuberculosis (MTB) complex than conventional acid-fast bacilli (AFB) smears and MTB cultures. Using the recently developed AdvanSure TB/NTM kit (LG Life Science Diagnostic Division, Korea), which could differentiate nontuberculous mycobacteria (NTM) from MTB, this study compared clinical usefulness of real-time PCR assay and Amplicor MTB PCR assay (Roche Molecular Systems, USA) for diagnosis of tuberculosis.

Methods

A total of 213 specimens (148 respiratory and 65 nonrespiratory specimens) were tested by using real-time PCR, Amplicor MTB PCR, AFB smear, and MTB culture. The sensitivity and specificity of four methods were evaluated according to clinical diagnosis.

Results

Of six NTM grown in culture, four (67%) were detected by real-time PCR. The overall agreement of real-time and Amplicor MTB PCR was 92% (191/207). The overall sensitivity and specificity were 91% and 87%, respectively, for real-time PCR, and 86% and 93% for Amplicor MTB PCR. In nonrespi-ratory specimens, the sensitivities of real-time PCR, Amplicor MTB PCR, AFB smear, and MTB culture were 67%, 60%, 13%, and 40%, respectively, and the specificity of the four methods were all 100%.

Conclusion

For diagnosis of tuberculosis, the sensitivity and specificity of the real-time PCR assay using AdvanSure TB/NTM kit and Amplicor MTB PCR were similar, and the former could differentiate NTM from MTB. The PCR assay can be considered as a more sensitive technique for the detection of MTB than the conventional AFB smear and culture.

Keywords: Mycobacterium tuberculosis, Nontuberculous mycobacteria, nonrespiratory, Realtime PCR

References

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Table 1.

Results of 6 cases of nontuberculous mycobacteria (NTM)

No. Case	Specimen	Real-time PCR	Amplicor MTB PCR	AFB smear	Culture
1	Sputum	NTM	Neg	1＋	NTM∗
2	Sputum	NTM	Neg	3＋	NTM∗
3	Urine	NTM	Neg	3＋	M. avium
4	Sputum	NTM	Neg	Neg	M. abscessus
5	Sputum	Neg	Neg	Neg	M. fortuitum
6	Bronchial	Neg	Neg	Neg	M. intracellulare
	aspirate

^∗ The identification tests were not performed. Abbreviations: MTB, Mycobacterium tuberculosis; AFB, acid-fast bacilli; Neg, negative.

Table 2.

Comparison of results between real-time PCR and Amplicor MTB PCR assays for detection of MTB

		Real-time PCR
		TB (＋)	TB (−)	Total
Amplicor MTB PCR	TB (+) TB (−) Total	88∗ 13 101	3 103∗ 106	91 116 207

^∗ Concordance rate between real-time PCR and Amplicor MTB PCR was 92% (191/207). Abbreviations: MTB, Mycobacterium tuberculosis; TB, tuberculosis.

Table 3.

Sensitivity, specificity, predictive values of real-time PCR, Amplicor MTB PCR, AFB smear and culture for detection of MTB in all 207 specimens according to clinical diagnosis

	Sensitivity (%)	Specificity (%)	P value∗	Predictive Positive	value (%) Negative
Real-time PCR	91	87		86	92
Amplicor MTB PCR	86	93	0.324	91	89
AFB smear	46	100	0.000	100	68
Culture	59	100	0.000	100	74

^∗ Statistical analysis between real-time PCR and other methods (Amplicor MTB PCR, AFB, and culture) by chi-square test. Abbreviations: MTB, Mycobacterium tuberculosis; AFB, acid-fast bacilli.

Table 4.

Sensitivity and specificity of real-time PCR, Amplicor MTB PCR, AFB smear and culture for detection of MTB according to respiratory and nonrespiratory specimens

	Sensitivity (%)		Specificity (%)		P value∗
	Resp	NonR	Resp	NonR	Resp	NonR
Real-time PCR	95	67	77	100
Amplicor MTB PCR	91	60	87	100	0.276	0.804
AFB smear	52	13	100	100	0.000	0.015
Culture	63	40	100	100	0.000	0.285

^∗ Statistical analysis between real-time PCR and other methods (Amplicor MTB PCR, AFB smear, and culture) by chi-square test. Abbreviations: MTB, Mycobacterium tuberculosis; AFB, acid-fast bacilli; Resp, respiratory specimens; NonR, nonrespiratory specimens.

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