Development and Efficacy of Real-Time PCR in the Diagnosis of Vivax Malaria Using Field Samples in the Republic of Korea

Jung-Yeon Kim,Dong-Il Chung,Youn-Kyoung Goo,Won-Ja Lee,So-Young Ji,Shin-Hyung Cho,Eun-Taek Han,Yeonchul Hong,Hyun-Il Shin,Takafumi Tsuboi

doi:10.1371/journal.pone.0105871

Abstract

The development of sensitive, rapid, and accurate diagnostic methods for vivax malaria is essential for the effective control of malaria in the Republic of Korea, where vivax malaria patients usually show low parasitemia. In this study, a TaqMan-based real-time polymerase chain reaction (PCR) method was established and compared with other PCR-based assays, including nested PCR, loop-mediated isothermal amplification, and multiplex PCR, using samples from febrile patients with suspected vivax malaria. The established real-time PCR had a high sensitivity (99.6%) and specificity (100%). Therefore, this sensitive, specific, rapid, and quantitative real-time PCR method could be used for the routine diagnosis of vivax malaria in the laboratory of the Korea National Institute of Health.

Highlights

Malaria caused by Plasmodium spp. occurs worldwide
Microscopic examination Microscopic examination for the limit of detection (LOD) calculation was performed on blood samples collected from vivax malaria patients whose infection had been previously confirmed by nested polymerase chain reaction (PCR) and microscopic examination
Vivax malaria in the Republic of Korea (ROK) is characterized by low parasitemia and a long latency period after primary infection in humans

Summary

Introduction

Malaria caused by Plasmodium spp. occurs worldwide. According to the World Malaria Report published by the World Health Organization (WHO), 207 million malaria cases occurred among 3.3 billion at-risk people from 103 countries in 2012, resulting in an estimated 627,000 deaths [1]. To date, nested PCR, loop-mediated isothermal amplification (LAMP), and real-time PCR methods have been developed to detect vivax malaria [14,23,24]. The present study aimed to establish a TaqMan-based real-time PCR method and compare its limit of detection (LOD) with other PCR-based assays, i.e., nested PCR, LAMP, and multiplex PCR.

Results

Conclusion