Molecular test for vivax malaria with loop-mediated isothermal amplification method in central China

Abstract

Vivax malaria has the highest incidence in central China. High-throughput and cost-effective testing methods are essential for vivax malaria diagnosis in this area. Loop-mediated isothermal amplification (LAMP) is an established nucleic acid amplification method, which provides a promising platform for the molecular detection of malaria parasites in development countries. This study was performed to compare the LAMP method, the nested PCR-based method, and microscopic examinations in diagnosing vivax malaria. LAMP detected vivax malaria parasites in 160 of 164 microscopically positive samples (sensitivity, 97.6%), whereas nested PCR detected Plasmodium vivax in 162 of 164 samples (sensitivity, 98.8%). No false-positive results were obtained by LAMP or nested PCR among fever-positive and healthy samples. The sensitivities and specificities of the two molecular tests were consistently high. In addition, the reproducibility of the LAMP assays using water bath, which reduced the cost substantially. LAMP method is an accurate, rapid, simple, and cost-effective method that may be useful for diagnosis in field diagnoses instead of nested PCR. This method is feasible to diagnose vivax malaria parasite in endemic areas of central China.

Rapid detection of Plasmodium falciparum with isothermal recombinase polymerase amplification and lateral flow analysis

“…Examples are the loop-mediated isothermal amplification (LAMP) for the detection of P. falciparum [6], Plasmodium vivax [7] or for four species of human malaria parasites [8] and the real-time quantitative nucleic acid sequence based amplification (real-time QT-NASBA) for the quantification of rRNA samples of three different Plasmodium strains [9]. These methods drastically simplified the assay set-up and improved the speed for nucleic acid amplification in comparison to a standard PCR.…”

Diagnosis and Treatment of Plasmodium vivax Malaria

“…A modified LAMP procedure, as well as standard nested PCR, was performed in the diagnosis of P. vivax in large field surveys in China. 19 Serology. Diagnostic techniques employing serological markers have been applied mostly to studies of protective immunity and epidemiology.…”

Prevalence of Drug Resistance-Associated Gene Mutations in Plasmodium vivax in Central China

“…Ethical approval was obtained from the ethical review committees at the National Institute of Parasitic Diseases, Chinese CDC, and the Walter Reed Army Institute of Research, USA. Filter papers with whole blood deposits were used for DNA elution by methods described previously [16]. To avoid cross-contamination, punch was cleaned in 70% ethanol and then punched a clean filter paper 3 times before cutting a new sample.…”