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RESEARCH ARTICLE
Year : 2017  |  Volume : 54  |  Issue : 1  |  Page : 54-60

Evaluation of SYBR green I based visual loop-mediated isothermal amplification (LAMP) assay for genus and species-specific diagnosis of malaria in P. vivax and P. falciparum endemic regions


1 National Institute of Malaria Research (ICMR), Dwarka; National Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi, India
2 National Institute of Malaria Research (ICMR), Dwarka, India
3 National Institute of Malaria Research, Field Unit, Raipur, Chhattisgarh, India

Correspondence Address:
Ruchi Singh
Scientist ‘E’, National Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi-110 029
India
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Source of Support: None, Conflict of Interest: None


PMID: 28352046

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Background & objectives: Loop-mediated isothermal amplification (LAMP) is an emerging nucleic acid based diagnostic approach that is easily adaptable to the field settings with limited technical resources. This study was aimed to evaluate the LAMP assay for the detection and identification of Plasmodium falciparum and P. vivax infection in malaria suspected cases using genus and species-specific assay. Methods: The 18S rRNA-based LAMP assay was evaluated for diagnosis of genus Plasmodium, and species-specific diagnosis of P. falciparum and P. vivax, infection employing 317 malaria suspected cases, and the results were compared with those obtained by 18S nested PCR (n-PCR). All the samples were confirmed by microscopy for the presence of Plasmodium parasite. Results: The n-PCR was positive in all Plasmodium-infected cases (n=257; P. falciparum=133; P. vivax=124) and negative in microscopy negative cases (n=58) except for two cases which were positive for P. vivax, giving a sensitivity of 100% (95% CI: 97.04-100%) and a specificity of 100% (95% CI: 88.45-99.5%). Genus-specific LAMP assay missed 11 (3.2%) microscopy and n-PCR confirmed vivax malaria cases. Considering PCR results as a reference, LAMP was 100% sensitive and specific for P. falciparum, whereas it exhibited 95.16% sensitivity and 96.7% specificity for P. vivax. The n-PCR assay detected 10 mixed infection cases while species-specific LAMP detected five mixed infection cases of P. vivax and P. falciparum, which were not detected by microscopy. Interpretation & conclusion: Genus-specific LAMP assay displayed low sensitivity. Falciparum specific LAMP assay displayed high sensitivity whereas vivax specific LAMP assay displayed low sensitivity. Failed detection of vivax cases otherwise confirmed by the n-PCR assay indicates exploitation of new targets and improved detection methods to attain 100% results for P. vivax detection.


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