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Recombinant ligA for leptospirosis diagnosis and ligA among the Leptospira spp. clinical isolates.
J Microbiol Methods. 2008 Jan; 72(1):73-81.JM

Abstract

Rapid diagnosis for differentiation of leptospirosis from other pyrogenic infections prevailing in the same locality is imperative for proper treatment. During infection, the pathogenic Leptospira spp. express virulence factors which induce antibody responses in the infected host. In this study, 50 referenced Leptospira spp. belonging to six genomospecies and 10 L. interrogans clinical isolates were studied for the presence of a gene encoding an in vivo expressed, surface exposed, immunoglobulin-like protein, LigA, by using PCR and southern hybridization specific to the 5' terminus sequence of the DNA. LigA was also detected in the Leptospira spp. whole cell homogenates by a direct ELISA using a mouse antiserum to the C-terminal portion of recombinant LigA (cLigA) as a detection reagent. All pathogenic Leptospira spp. except one of the two strains of L. santorasai were positive for the gene and its phenotype while all of the L. borgpetersenii and L. biflexa strains were negative. Recombinant cLigA was used as an antigen in ELISAs for detecting IgM and IgG in the sera of leptospirosis patients and in the sera of patients with other febrile illnesses and healthy subjects. When acute phase sera were tested by the cLigA IgM- and IgG-ELISAs, 92% and 100% of the MAT-positive sera were positive, respectively. The diagnostic sensitivity was 100% when both IgM- and IgG-ELISAs were performed on the same acute phase sera and the results were combined. Acute and convalescence sera of patients who were Leptospira culture positive but MAT/IgM-dipstick negative gave 88% and 100% positives by combined cLigA IgM/IgG ELISAs. The diagnostic specificities for the cLigA IgM- and IgG-ELISAs were 98% and 100%, respectively. Our cLigA based-serology has a high potential for early diagnosis of leptospirosis especially when the culture and MAT results are not yet available.

Authors+Show Affiliations

Faculty of Allied Health Sciences, Thammasat University, Rangsit Campus, Pathumthani, Thailand. psrimanote01@yahoo.com.auNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

18079011

Citation

Srimanote, Potjanee, et al. "Recombinant ligA for Leptospirosis Diagnosis and ligA Among the Leptospira Spp. Clinical Isolates." Journal of Microbiological Methods, vol. 72, no. 1, 2008, pp. 73-81.
Srimanote P, Wongdeethai N, Jieanampunkul P, et al. Recombinant ligA for leptospirosis diagnosis and ligA among the Leptospira spp. clinical isolates. J Microbiol Methods. 2008;72(1):73-81.
Srimanote, P., Wongdeethai, N., Jieanampunkul, P., Samonkiert, S., Leepiyasakulchai, C., Kalambaheti, T., & Prachayasittikul, V. (2008). Recombinant ligA for leptospirosis diagnosis and ligA among the Leptospira spp. clinical isolates. Journal of Microbiological Methods, 72(1), 73-81.
Srimanote P, et al. Recombinant ligA for Leptospirosis Diagnosis and ligA Among the Leptospira Spp. Clinical Isolates. J Microbiol Methods. 2008;72(1):73-81. PubMed PMID: 18079011.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Recombinant ligA for leptospirosis diagnosis and ligA among the Leptospira spp. clinical isolates. AU - Srimanote,Potjanee, AU - Wongdeethai,Nattawut, AU - Jieanampunkul,Pathumporn, AU - Samonkiert,Suwanna, AU - Leepiyasakulchai,Chaniya, AU - Kalambaheti,Thareerat, AU - Prachayasittikul,Virapong, Y1 - 2007/11/17/ PY - 2007/09/11/received PY - 2007/10/29/revised PY - 2007/10/31/accepted PY - 2007/12/15/pubmed PY - 2008/3/26/medline PY - 2007/12/15/entrez SP - 73 EP - 81 JF - Journal of microbiological methods JO - J Microbiol Methods VL - 72 IS - 1 N2 - Rapid diagnosis for differentiation of leptospirosis from other pyrogenic infections prevailing in the same locality is imperative for proper treatment. During infection, the pathogenic Leptospira spp. express virulence factors which induce antibody responses in the infected host. In this study, 50 referenced Leptospira spp. belonging to six genomospecies and 10 L. interrogans clinical isolates were studied for the presence of a gene encoding an in vivo expressed, surface exposed, immunoglobulin-like protein, LigA, by using PCR and southern hybridization specific to the 5' terminus sequence of the DNA. LigA was also detected in the Leptospira spp. whole cell homogenates by a direct ELISA using a mouse antiserum to the C-terminal portion of recombinant LigA (cLigA) as a detection reagent. All pathogenic Leptospira spp. except one of the two strains of L. santorasai were positive for the gene and its phenotype while all of the L. borgpetersenii and L. biflexa strains were negative. Recombinant cLigA was used as an antigen in ELISAs for detecting IgM and IgG in the sera of leptospirosis patients and in the sera of patients with other febrile illnesses and healthy subjects. When acute phase sera were tested by the cLigA IgM- and IgG-ELISAs, 92% and 100% of the MAT-positive sera were positive, respectively. The diagnostic sensitivity was 100% when both IgM- and IgG-ELISAs were performed on the same acute phase sera and the results were combined. Acute and convalescence sera of patients who were Leptospira culture positive but MAT/IgM-dipstick negative gave 88% and 100% positives by combined cLigA IgM/IgG ELISAs. The diagnostic specificities for the cLigA IgM- and IgG-ELISAs were 98% and 100%, respectively. Our cLigA based-serology has a high potential for early diagnosis of leptospirosis especially when the culture and MAT results are not yet available. SN - 0167-7012 UR - https://www.unboundmedicine.com/medline/citation/18079011/Recombinant_ligA_for_leptospirosis_diagnosis_and_ligA_among_the_Leptospira_spp__clinical_isolates_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0167-7012(07)00354-5 DB - PRIME DP - Unbound Medicine ER -