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Loop-mediated isothermal amplification method targeting the TTS1 gene cluster for detection of Burkholderia pseudomallei and diagnosis of melioidosis.
J Clin Microbiol. 2008 Feb; 46(2):568-73.JC

Abstract

Melioidosis is a severe infection caused by Burkholderia pseudomallei. The timely implementation of effective antimicrobial treatment requires rapid diagnosis. Loop-mediated isothermal amplification (LAMP) targeting the TTS1 gene cluster was developed for the detection of B. pseudomallei. LAMP was sensitive and specific for the laboratory detection of this organism. The lower limit of detection was 38 genomic copies per reaction, and LAMP was positive for 10 clinical B. pseudomallei isolates but negative for 5 B. thailandensis and 5 B. mallei isolates. A clinical evaluation was conducted in northeast Thailand to compare LAMP to an established real-time PCR assay targeting the same TTS1 gene cluster. A total of 846 samples were obtained from 383 patients with suspected melioidosis, 77 of whom were subsequently diagnosed with culture-confirmed melioidosis. Of these 77 patients, a positive result was obtained from one or more specimens by PCR in 26 cases (sensitivity, 34%; 95% confidence interval [CI], 23.4 to 45.4%) and by LAMP in 34 cases (sensitivity, 44%; 95% CI, 32.8 to 55.9%) (P = 0.02). All samples from 306 patients that were culture negative for B. pseudomallei were negative by PCR (specificity, 100%; 95% CI, 98.8 to 100%), but 5 of 306 patients (1.6%) were positive by LAMP (specificity, 98.4%; 95% CI, 96.2 to 99.5%) (P = 0.03). The diagnostic accuracies of PCR and LAMP were 86.7% (95% CI, 82.9 to 89.9%) and 87.5% (95% CI, 83.7 to 90.6%), respectively (P = 0.47). Both assays were very insensitive when applied to blood samples; PCR and LAMP were positive for 0 and 1 of 44 positive blood cultures, respectively. The PCR and LAMP assays evaluated here are not sufficiently sensitive to replace culture in our clinical setting.

Authors+Show Affiliations

Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, 420/6 Rajvithi Road, Bangkok 10400, Thailand. narisara@tropmedres.acNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18039797

Citation

Chantratita, Narisara, et al. "Loop-mediated Isothermal Amplification Method Targeting the TTS1 Gene Cluster for Detection of Burkholderia Pseudomallei and Diagnosis of Melioidosis." Journal of Clinical Microbiology, vol. 46, no. 2, 2008, pp. 568-73.
Chantratita N, Meumann E, Thanwisai A, et al. Loop-mediated isothermal amplification method targeting the TTS1 gene cluster for detection of Burkholderia pseudomallei and diagnosis of melioidosis. J Clin Microbiol. 2008;46(2):568-73.
Chantratita, N., Meumann, E., Thanwisai, A., Limmathurotsakul, D., Wuthiekanun, V., Wannapasni, S., Tumapa, S., Day, N. P., & Peacock, S. J. (2008). Loop-mediated isothermal amplification method targeting the TTS1 gene cluster for detection of Burkholderia pseudomallei and diagnosis of melioidosis. Journal of Clinical Microbiology, 46(2), 568-73.
Chantratita N, et al. Loop-mediated Isothermal Amplification Method Targeting the TTS1 Gene Cluster for Detection of Burkholderia Pseudomallei and Diagnosis of Melioidosis. J Clin Microbiol. 2008;46(2):568-73. PubMed PMID: 18039797.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Loop-mediated isothermal amplification method targeting the TTS1 gene cluster for detection of Burkholderia pseudomallei and diagnosis of melioidosis. AU - Chantratita,Narisara, AU - Meumann,Ella, AU - Thanwisai,Aunchalee, AU - Limmathurotsakul,Direk, AU - Wuthiekanun,Vanaporn, AU - Wannapasni,Saran, AU - Tumapa,Sarinna, AU - Day,Nicholas P J, AU - Peacock,Sharon J, Y1 - 2007/11/26/ PY - 2007/11/28/pubmed PY - 2008/3/19/medline PY - 2007/11/28/entrez SP - 568 EP - 73 JF - Journal of clinical microbiology JO - J Clin Microbiol VL - 46 IS - 2 N2 - Melioidosis is a severe infection caused by Burkholderia pseudomallei. The timely implementation of effective antimicrobial treatment requires rapid diagnosis. Loop-mediated isothermal amplification (LAMP) targeting the TTS1 gene cluster was developed for the detection of B. pseudomallei. LAMP was sensitive and specific for the laboratory detection of this organism. The lower limit of detection was 38 genomic copies per reaction, and LAMP was positive for 10 clinical B. pseudomallei isolates but negative for 5 B. thailandensis and 5 B. mallei isolates. A clinical evaluation was conducted in northeast Thailand to compare LAMP to an established real-time PCR assay targeting the same TTS1 gene cluster. A total of 846 samples were obtained from 383 patients with suspected melioidosis, 77 of whom were subsequently diagnosed with culture-confirmed melioidosis. Of these 77 patients, a positive result was obtained from one or more specimens by PCR in 26 cases (sensitivity, 34%; 95% confidence interval [CI], 23.4 to 45.4%) and by LAMP in 34 cases (sensitivity, 44%; 95% CI, 32.8 to 55.9%) (P = 0.02). All samples from 306 patients that were culture negative for B. pseudomallei were negative by PCR (specificity, 100%; 95% CI, 98.8 to 100%), but 5 of 306 patients (1.6%) were positive by LAMP (specificity, 98.4%; 95% CI, 96.2 to 99.5%) (P = 0.03). The diagnostic accuracies of PCR and LAMP were 86.7% (95% CI, 82.9 to 89.9%) and 87.5% (95% CI, 83.7 to 90.6%), respectively (P = 0.47). Both assays were very insensitive when applied to blood samples; PCR and LAMP were positive for 0 and 1 of 44 positive blood cultures, respectively. The PCR and LAMP assays evaluated here are not sufficiently sensitive to replace culture in our clinical setting. SN - 1098-660X UR - https://www.unboundmedicine.com/medline/citation/18039797/Loop_mediated_isothermal_amplification_method_targeting_the_TTS1_gene_cluster_for_detection_of_Burkholderia_pseudomallei_and_diagnosis_of_melioidosis_ L2 - https://journals.asm.org/doi/10.1128/JCM.01817-07?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -