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Evaluation of a modified Taqman assay detecting pathogenic Leptospira spp. against culture and Leptospira-specific IgM enzyme-linked immunosorbent assay in a clinical environment.
Diagn Microbiol Infect Dis. 2007 Apr; 57(4):361-6.DM

Abstract

A Taqman assay for the detection of pathogenic Leptospira was modified to suit the LightCycler instrument. The modified assay was found to have an analytical sensitivity of 10 copies/reaction. The assay was then compared to the current gold standard for acute phase detection, culture, and to a commercially available Leptospira-specific IgM enzyme-linked immunosorbent assay (ELISA). Of the 236 samples including serum and ethylenediaminetetraacetic acid anticoagulated blood sample submitted for testing, polymerase chain reaction (PCR) was able to detect Leptospira DNA in 27 of the 28 culture positives and in 1 negative culture. Discrepant results were resolved by using the microscopic agglutination test on convalescent sera. The PCR was found to have a clinical specificity and sensitivity of 99.5% and 96.4%, respectively, when compared to the culture. Comparisons between culture and ELISA showed that the ELISA lack sensitivity (4.2%) and positive predictability (3.6%) for the detection of acute phase Leptospira infections. These results show that the modified LightCycler Taqman assay could be used as a replacement of culture for the detection of pathogenic Leptospira in a clinical setting.

Authors+Show Affiliations

WHO/FAO/OIE Collaborating Centre for Reference and Research on Leptospirosis, Western Pacific Region, Centre for Public Health Sciences, Queensland Health Scientific Services, Brisbane 4108, Australia. andrew_slack@health.qld.gov.auNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Evaluation Study
Journal Article

Language

eng

PubMed ID

17188447

Citation

Slack, Andrew, et al. "Evaluation of a Modified Taqman Assay Detecting Pathogenic Leptospira Spp. Against Culture and Leptospira-specific IgM Enzyme-linked Immunosorbent Assay in a Clinical Environment." Diagnostic Microbiology and Infectious Disease, vol. 57, no. 4, 2007, pp. 361-6.
Slack A, Symonds M, Dohnt M, et al. Evaluation of a modified Taqman assay detecting pathogenic Leptospira spp. against culture and Leptospira-specific IgM enzyme-linked immunosorbent assay in a clinical environment. Diagn Microbiol Infect Dis. 2007;57(4):361-6.
Slack, A., Symonds, M., Dohnt, M., Harris, C., Brookes, D., & Smythe, L. (2007). Evaluation of a modified Taqman assay detecting pathogenic Leptospira spp. against culture and Leptospira-specific IgM enzyme-linked immunosorbent assay in a clinical environment. Diagnostic Microbiology and Infectious Disease, 57(4), 361-6.
Slack A, et al. Evaluation of a Modified Taqman Assay Detecting Pathogenic Leptospira Spp. Against Culture and Leptospira-specific IgM Enzyme-linked Immunosorbent Assay in a Clinical Environment. Diagn Microbiol Infect Dis. 2007;57(4):361-6. PubMed PMID: 17188447.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Evaluation of a modified Taqman assay detecting pathogenic Leptospira spp. against culture and Leptospira-specific IgM enzyme-linked immunosorbent assay in a clinical environment. AU - Slack,Andrew, AU - Symonds,Meegan, AU - Dohnt,Michael, AU - Harris,Christopher, AU - Brookes,Dianne, AU - Smythe,Lee, Y1 - 2006/12/22/ PY - 2006/05/30/received PY - 2006/10/03/revised PY - 2006/10/04/accepted PY - 2006/12/26/pubmed PY - 2007/6/22/medline PY - 2006/12/26/entrez SP - 361 EP - 6 JF - Diagnostic microbiology and infectious disease JO - Diagn Microbiol Infect Dis VL - 57 IS - 4 N2 - A Taqman assay for the detection of pathogenic Leptospira was modified to suit the LightCycler instrument. The modified assay was found to have an analytical sensitivity of 10 copies/reaction. The assay was then compared to the current gold standard for acute phase detection, culture, and to a commercially available Leptospira-specific IgM enzyme-linked immunosorbent assay (ELISA). Of the 236 samples including serum and ethylenediaminetetraacetic acid anticoagulated blood sample submitted for testing, polymerase chain reaction (PCR) was able to detect Leptospira DNA in 27 of the 28 culture positives and in 1 negative culture. Discrepant results were resolved by using the microscopic agglutination test on convalescent sera. The PCR was found to have a clinical specificity and sensitivity of 99.5% and 96.4%, respectively, when compared to the culture. Comparisons between culture and ELISA showed that the ELISA lack sensitivity (4.2%) and positive predictability (3.6%) for the detection of acute phase Leptospira infections. These results show that the modified LightCycler Taqman assay could be used as a replacement of culture for the detection of pathogenic Leptospira in a clinical setting. SN - 0732-8893 UR - https://www.unboundmedicine.com/medline/citation/17188447/Evaluation_of_a_modified_Taqman_assay_detecting_pathogenic_Leptospira_spp__against_culture_and_Leptospira_specific_IgM_enzyme_linked_immunosorbent_assay_in_a_clinical_environment_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0732-8893(06)00384-1 DB - PRIME DP - Unbound Medicine ER -