Performance Evaluation of Anyplex plus MTB/NTM and AdvanSure TB/NTM for the Detection of Mycobacterium tuberculosis and Nontuberculous Mycobacteria

Wonho Choe; Ehwa Kim; Seo Yeon Park; Jeong Don Chae

doi:10.5145/ACM.2015.18.2.44

Journal List > Ann Clin Microbiol > v.18(2) > 1078534

Go to TopGo to Top Go to BottomGo to Bottom

TOOLS

Choe, Kim, Park, and Chae: Performance Evaluation of Anyplex plus MTB/NTM and AdvanSure TB/NTM for the Detection of Mycobacterium tuberculosis and Nontuberculous Mycobacteria

Original Article

Ann Clin Microbiol 2015;18(2):44-51.

Published online: 5 June 2015

DOI: https://doi.org/10.5145/ACM.2015.18.2.44

Performance Evaluation of Anyplex plus MTB/NTM and AdvanSure TB/NTM for the Detection of Mycobacterium tuberculosis and Nontuberculous Mycobacteria

Wonho Choe, Ehwa Kim, Seo Yeon Park, Jeong Don Chae

Department of Laboratory Medicine, Eulji General Hospital, Eulji University College of Medicine, Seoul, Korea

Correspondence: Jeong Don Chae, Department of Laboratory Medicine, Eulji General Hospital, Eulji University College of Medicine, 68, Hangeulbiseok-ro, Nowon-gu, Seoul 139-711, Korea. (Tel) 82-2-970-8324, (Fax) 82-2-970-8524, (E-mail) jdchae@eulji.ac.kr

Received 20 November 201426 May 2015 Accepted 12 June 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

초록

Background

Polymerase chain reaction (PCR) methods from direct specimen are widely used for the rapid and accurate detection of mycobacteria infection. In this study, we evaluated two domestically developed detection kits for Mycobacterium tuberculosis (MTB) and nontuberculous mycobacteria (NTM) using re-al-time PCR.

Methods

A total of 348 samples from patients with suspected tuberculosis were tested with real-time PCR over seven months. We performed real-time PCR using the recently developed Anyplex plus MTB/NTM Detection kit (Seegene) with the CFX 96TM Real- time PCR System (Bio-Rad Laboratories) and the conventional AdvanSure TB/NTM real-time PCR kit (LG Life Sciences) with the SLAN Real-time PCR detection system (LG Life Sciences) to evaluate their performance for detecting MTB and NTM.

Results

The two real-time PCR systems showed 96.8% concordance rate for MTB-positive, NTM-positive, and negative results. Based on culture results, the sensitivity and specificity for the detection of MTB using PCR were 71.0% and 94.9% for Anyplex plus, and 78.1% and 93.9% for AdvanSure, respectively. For the detection of NTM, the sensitivity and specificity were 33.3% and 98.4% for Anyplex plus, and 51.7% and 97.9% for AdvanSure. Both PCR systems showed high MTB positive results in bronchial wash-ing and sputum samples.

Conclusion

In detecting MTB and NTM, Anyplex plus MTB/NTM (Seegene) and AdvanSure TB/NTM real-ti-me PCR (LG Life Sciences) showed high concordance rate with each other in all samples. Therefore both detection kits can be used as rapid and reliable detection tool for MTB.

Keywords: Nontuberculous mycobacteria, Real-time PCR, Tuberculosis

REFERENCES

1.Joint Committee for the Revision of Korean Guidelines for Tuberculosis Korea Centers for Disease Control and Prevention. Korean Guidelines for Tuberculosis. 2nd ed.Korea Centers for Disease Control and Prevention;2014. p. 30–1.

2.Jang MA., Shin SY., Park S., Seong MW., Park SS., Kim SH. Diagnostic Genetics Subcommittee, The Korean Association of Quality Assurance for Clinical Laboratory. Annual report on the external quality assessment of diagnostic genetics in Korea (2013). J Lab Med Qual Assur. 2014. 36:71–83.

3.Korea Centers for Disease Control and Prevention. Manual of Laboratory Tests for Tuberculosis. Korea Centers for Disease Control and Prevention. 2013. 75–80.

4.Joint Committee for the Revision of Korean Guidelines for Tuberculosis Korea Centers for Disease Control and Prevention. Korean Guidelines for Tuberculosis. 2nd ed.Korea Centers for Disease Control and Prevention;2014. p. 227–8.

5.Shinnick TM., Iademarco MF., Ridderhof JC. National plan for reliable tuberculosis laboratory services using a systems approach. Recommendations from CDC and the Association of Public Health Laboratories Task Force on Tuberculosis Laboratory Services.

6.Centers for Disease Control and Prevention (CDC). Updated guidelines for the use of nucleic acid amplification tests in the diagnosis of tuberculosis. MMWR Morb Mortal Wkly Rep. 2009. 58:7–10.

7.Broccolo F., Scarpellini P., Locatelli G., Zingale A., Brambilla AM., Cichero P, et al. Rapid diagnosis of mycobacterial infections and quantitation of Mycobacterium tuberculosis load by two real-time calibrated PCR assays. J Clin Microbiol. 2003. 41:4565–72.

8.Chang HE., Heo SR., Yoo KC., Song SH., Kim SH., Kim HB, et al. Detection of Mycobacterium tuberculosis complex using real-time polymerase chain reaction. Korean J Lab Med. 2008. 28:103–8.

9.Lim J., Kim J., Kim JW., Ihm C., Sohn YH., Cho HJ, et al. Multicenter evaluation of Seegene Anyplex TB PCR for the detection of Mycobacterium tuberculosis in respiratory specimens. J Microbiol Biotechnol. 2014. 24:1004–7.

10.Lee JH., Kim BH., Lee MK. Performance evaluation of anyplex plus MTB/NTM and MDR-TB detection kit for detection of mycobacteria and for antituberculosis drug susceptibility test. Ann Clin Microbiol. 2014. 17:115–22.

11.Lee H., Park KG., Lee G., Park J., Park YG., Park YJ. Assessment of the quantitative ability of AdvanSure TB/NTM real-time PCR in respiratory specimens by comparison with phenotypic methods. Ann Lab Med. 2014. 34:51–5.

12.Cho SY., Kim MJ., Suh JT., Lee HJ. Comparison of diagnostic performance of three real-time PCR kits for detecting Mycobacterium species. Yonsei Med J. 2011. 52:301–6.

Fig. 1.

Type distributions of samples according to the tests performed. Abbreviations: BAL, bronchoalveolar lavage; Pleural, pleural fluid; CSF, cerebrospinal fluid, SM, solid media; LM, liquid media.

Table 1.

Results of two real-time PCR systems according to AFB smear and culture results

Target	AFB smear	Assays	Culture + (79)		Culture − (200)		Sensitivity/Specificity	PPV/NPV
Target	AFB smear	Assays	PCR+	PCR−	PCR+	PCR−	Sensitivity/Specificity	PPV/NPV
MTB	Positive	Anyplex plus	23	1	0	2	95.8/100.0	100.0/66.7
		AdvanSure	23	0	0	2	100.0/100.0	100.0/100.0
	Negative	Anyplex plus	25	15	10	185	62.5/94.9	71.4/92.5
		AdvanSure	25	11	12	182	69.4/93.8	67.6/94.3
	All∗	Anyplex plus	49†	20§	10	187	71.0/94.9	83.1/90.3
		AdvanSure	50‡	14∥	12	184	78.1/93.9	80.6/92.9
NTM	Positive	Anyplex plus	9	1	0	2	90.0/100.0	100.0/66.7
		AdvanSure	10	0	0	2	100.0/100.0	100.0/100.0
	Negative	Anyplex plus	1	15	3	185	6.3/98.4	25.0/92.5
		AdvanSure	5	11	4	182	31.3/97.8	55.6/94.3
	All∗	Anyplex plus AdvanSure	1015	20§14∥	3 4	187184	33.3/98.4 51.7/97.9	76.9/90.3 78.9/92.9

^∗ Includes samples without AFB smear results:

^† n=1;

^‡ n=2;

^§ n=4;

^∥ n=3.

Abbreviations: MTB, mycobacterium tuberculosis; NTM, nontuberculous mycobacteria; AdvanSure, AdvanSure TB/NTM real-time PCR; Anyplex plus, Anyplex plus MTB/NTM Detection; AFB, acid-fast bacilli; PCR, polymerase chain reaction; PPV, positive predictive value; NPV, negative predictive value.

Table 2.

MTB positive rates according to sample types in culture and two real-time PCR systems

Specimen	No. MTB positive (%)
Specimen	Culture	Anyplex plus	AdvanSure
Pulmonary specimen	53/267 (19.9%)	58/267 (21.7%)	60/267 (22.5%)
Extrapulmonary specimen∗	4/12 (33.3%)	1/12 (8.3%)	2/12 (16.7%)
Total	57/279 (20.4%)	59/279 (22.1%)	65/279 (22.2%)

^∗ CSF (7), Open pus (3), Colon (1), Tissue (1).

Abbreviations: AdvanSure, AdvanSure TB/NTM real-time PCR; Anyplex plus, Anyplex plus MTB/NTM Detection; MTB, Mycobacterium tuberculosis.

Table 3.

Results of two real-time PCR systems according to culture results

	Anyplex plus PCR	AdvanSure PCR	Number
Culture + (79)	+	+	59
	+	−	0
	−	+	6
	−	−	14
Culture − (200)	+	+	13
	+	−	0
	−	+	3
	−	−	184

Abbreviations: AdvanSure, AdvanSure TB/NTM real-time PCR;

Anyplex plus, Anyplex plus MTB/NTM Detection.

Table 4.

Result correlations of two real-time PCR systems

		Anyplex plus
		MTB	NTM	Negative	Total
AdvanSure	MTB	61	0	4	65
	NTM	1	13	5	19
	Negative	1	0	256	257
	Total	63	13	265	341

Abbreviations: AdvanSure, AdvanSure TB/NTM real-time PCR; Anyplex plus, Anyplex plus MTB/NTM Detection; MTB, Mycobacterium tuberculosis; NTM, nontuberculous mycobacteria.

Table 5.

Result summary for samples with discrepant results among test methods including culture, AFB smear, and PCR

No	Sample	AdvanSure	Ct	Anyplex plus	Ct	Culture	Culture media	NTM identification	AFB smear	Treatment
1	BAL	MTB	33.5	-		-			−	Done
2	BAL	MTB	33.5	-		-			−	Done
3	Sputum	MTB	34.5	-		-			−
4	Colon	MTB	34.0	-		MTB	SM 1		NT	Done
5	BAL	NTM	31.8	MTB	42.2	NTM	SM 1, LM 1	M. avium	−	Done
6	Pleural fluid	-		MTB	43.6	NT			−	Done
7	BAL	NTM	33.8	-	37.2	NTM	SM 2, LM 2	M. intracellulare	−
8	Sputum	NTM	25.6	-		NTM	SM 1, LM 1	M. intracellulare	+ (few)
9	Sputum	NTM	23.6	-		NTM	SM 2, LM 2	M. intracellulare	−
10	BAL	NTM	32.6	-	35.4	NTM	SM 1, LM 1	NT	−
11	Sputum	NTM	33.9	-	36.5	-			−

Abbreviations: AdvanSure, AdvanSure TB/NTM real-time PCR; Anyplex plus, Anyplex plus MTB/NTM Detection; AFB, acid-fast bacilli; BAL, bronchoalveolar lavage; Ct, cycle threshold; MTB, Mycobacterium tuberculosis; NTM, nontuberculous mycobacteria; SM, solid media; LM, liquid media; NT, not tested.

TOOLS

Similar articles