New molecular detection methods of malaria parasites with multiplegenes from genomes

Gupta, Himanshu and Srivastava, Shikha and Chaudhari, Sima and Vasudevan, Thanvanthri G and Hande, Manjunath H and Umakanth, Shashikiran and Satyamoorthy, K (2016) New molecular detection methods of malaria parasites with multiplegenes from genomes. Acta Tropica, 160. pp. 15-22. ISSN 0001-706X

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Abstract

For the effective control of malaria, development of sensitive, accurate and rapid tool to diagnose andmanage the disease is essential. In humans subjects, the severe form of malaria is caused by Plasmodiumfalciparum (Pf) and Plasmodium vivax (Pv) and there is need to identify these parasites in acute, chronicand latent (during and post-infection) stages of the disease. In this study, we report a species specificand sensitive diagnostic method for the detection of Pf and Pv in humans. First, we identified intra andintergenic multiloci short stretch of 152 (PfMLS152) and 110 (PvMLS110) nucleotides which is present upto 44 and 34 times in the genomes of Pf and Pv respectively. We developed the single-step amplification-based method using isolated DNA or from lysed red blood cells for the detection of the two malariaparasites. The limit of detection of real-time polymerase chain reaction based assays were 0.1copyofparasite/�l for PfMLS152 and PvMLS110 target sequences. Next, we have tested 250 clinically suspectedcases of malaria to validate the method. Sensitivity and specificity for both targets were 100% compared tothe quantitative buffy coat microscopy analysis and real-time PCR (Pf-chloroquine resistance transporter(PfCRT) and Pv-lactate dehydrogenase (PvLDH)) based assays. The sensitivity of microscopy and real-timePCR (PfCRT and PvLDH primers) assays were 80.63%; 95%CI 75.22%–85.31%; p < 0.05 and 97.61%; 95%CI94.50%–99.21%; p < 0.05 in detecting malaria infection respectively when compared to PfMLS152 andPvMLS110 targets to identify malaria infection in patients. These improved assays may have potentialapplications in evaluating malaria in asymptomatic patients, treatment, blood donors and in vaccinestudies.

Item Type: Article
Uncontrolled Keywords: Malaria; repeat sequence; Real-time PCR; diagnosis; Multi-copy DNA.
Subjects: Life Sciences > MLSC Manipal
Medicine > KMC Manipal > Medicine
Depositing User: KMC Library
Date Deposited: 18 Aug 2016 09:48
Last Modified: 18 Aug 2016 09:48
URI: http://eprints.manipal.edu/id/eprint/146800

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