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Utility of Loop-mediated Isothermal Amplification Assay, Polymerase Chain Reaction, and ELISA for Diagnosis of Leptospirosis in South Indian Patients.

ABSTRACT: BACKGROUND:Leptospirosis is a zoonotic disease which requires laboratory diagnosis for confirmation. MATERIALS AND METHODS:In this study serum samples from adults with acute undifferentiated fever (duration ?15 days) were tested for IgM antibodies to Leptospira by ELISA, PCR for rrs gene and loop-mediated isothermal amplification (LAMP) assay for LipL32 and LipL41. RESULTS:Among the 150 sera tested, three were positive by PCR, LAMP and IgM ELISA/modified Faines' criteria, two by only PCR; seven only by LAMP assay and forty fulfilled modified Faine's criteria (illness clinically compatible and IgM ELISA positive for leptospirosis). Clinical correlation revealed renal compromise, low platelet count and severe jaundice were significantly related to leptospirosis (P < 0.05). CONCLUSION:This study suggests that LAMP assay could be useful for diagnosis of leptospirosis during the 1st week of illness whereas IgM ELISA forms the mainstay of diagnosis from the 2nd week onward. Further studies especially community based, comparing ELISA, PCR, LAMP, culture and microscopic agglutination test are required to evaluate the veracity of these findings.

SUBMITTER: Sengupta M

PROVIDER: S-EPMC5330041 | biostudies-literature | 2017 Jan-Mar

REPOSITORIES: biostudies-literature

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Utility of Loop-mediated Isothermal Amplification Assay, Polymerase Chain Reaction, and ELISA for Diagnosis of Leptospirosis in South Indian Patients.

Sengupta Mallika M Prabhakar Abhilash Kundavaram Paul AK Satyendra Sowmya S Thambu David D Abraham Ooriapadickal Cherian OC Balaji Veeraraghavan V Chen Hua-Wei HW Chao Chien-Chung CC Ching Wei-Mei WM Prakash John Antony Jude JA

Journal of global infectious diseases 20170101 1

<h4>Background</h4>Leptospirosis is a zoonotic disease which requires laboratory diagnosis for confirmation.<h4>Materials and methods</h4>In this study serum samples from adults with acute undifferentiated fever (duration ≤15 days) were tested for IgM antibodies to <i>Leptospira</i> by ELISA, PCR for <i>rrs</i> gene and loop-mediated isothermal amplification (LAMP) assay for LipL32 and LipL41.<h4>Results</h4>Among the 150 sera tested, three were positive by PCR, LAMP and IgM ELISA/modified Faine ...[more]

PMID: 28250618

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Project description:BackgroundOcular infection with Toxoplasma gondii is a major preventable cause of blindness, especially in young people. The aim of the present study was to assess detection rate of T. gondii DNA in blood samples of clinically diagnosed of ocular toxoplasmosis using uracil DNA glycosylase-supplemented loop-mediated isothermal amplification (UDG-LAMP) and real-time quantitative PCR (qPCR) based on REP-529 and B1.MethodsOne hundred and seventeen patients with clinically diagnosed ocular toxoplasmosis (OT) were participated in the study as well as 200 control patients. Peripheral blood samples were assessed using UDG-LAMP and qPCR techniques targeting REP-529 and B1.ResultsDetection limits of qPCR using REP-529 and B1 were estimated as 0.1 and 1 fg of T. gondii genomic DNA, respectively. The limits of detection for UDG-LAMP using REP-529 and B1 were 1 and 100 fg, respectively. In this study, 18 and 16 patients were positive in qPCR using REP-529 and B1, respectively. Based on the results of UDG-LAMP, 15 and 14 patients were positive using REP-529 and B1, respectively. Results of the study on patients with active ocular lesion showed that sensitivity of REP-529 and BI targets included 64 and 63%, respectively using qPCR. Sensitivity of 62 and 61%, were concluded from UDG-LAMP using REP-529 and B1 in the blood cases of active ocular lesion. qPCR was more sensitive than UDG-LAMP for the detection of Toxoplasma gondii DNA in peripheral blood samples of patients with clinically diagnosed toxoplasmic chorioretinitis. Furthermore, the REP-529 included a better detection rate for the diagnosis of ocular toxoplasmosis in blood samples, compared to that the B1 gene did. Moreover, the qPCR and UDG-LAMP specificity assessments have demonstrated no amplifications of DNAs extracted from other microorganisms based on REP-529 and B1.ConclusionsData from the current study suggest that qPCR and UDG-LAMP based on the REP-529 are promising diagnostic methods for the diagnosis of ocular toxoplasmosis in blood samples of patients with active chorioretinal lesions.

Dataset Information

Utility of Loop-mediated Isothermal Amplification Assay, Polymerase Chain Reaction, and ELISA for Diagnosis of Leptospirosis in South Indian Patients.

Publications

Utility of Loop-mediated Isothermal Amplification Assay, Polymerase Chain Reaction, and ELISA for Diagnosis of Leptospirosis in South Indian Patients.

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