Implementation of a novel PCR based method for detecting malaria parasites from naturally infected mosquitoes in Papua New Guinea

• Published in: Malaria Journal
• Main Authors: Amakawa Masao, Fujimoto Chigusa, Sattabongkot Jetsumon, Suguri Setsuo, Hasan Arif U, Harada Masakazu, Ohmae Hiroshi
• Format: Article in Journal/Newspaper
• Language: English
• Published: BMC 2009
• Subjects: Arctic medicine. Tropical medicine; RC955-962; Infectious and parasitic diseases; RC109-216; Arctic
• Online Access: https://doi.org/10.1186/1475-2875-8-182; https://doaj.org/article/a2e6e4e5402946309b19acabd35c6703

Abstract Background Detection of Plasmodium species in mosquitoes is important for designing vector control studies. However, most of the PCR-based detection methods show some potential limitations. The objective of this study was to introduce an effective PCR-based method for detecting Plasmodium vivax and Plasmodium falciparum from the field-caught mosquitoes of Papua New Guinea. Methods A method has been developed to concurrently detect mitochondrial cytochrome b ( Cyt b ) of four human Plasmodium species using PCR ( Cytb -PCR). To particularly discriminate P. falciparum from P. vivax , Plasmodium ovale and Plasmodium malariae , a polymerase chain reaction-repeated fragment length polymorphism (PCR-RFLP) has further been developed to use with this method. However, due to limited samples number of P. ovale and P. malariae this study was mainly confined to P. vivax and P. falciparum . The efficiency of Cytb -PCR was evaluated by comparing it with two 'gold standards' enzyme linked immunosorbent assay specific for circumsporozoite protein (CS-ELISA) using artificially infected mosquitoes; and nested PCR specific for small subunit ribosomal RNA ( SSUrRNA ) using field caught mosquitoes collected from three areas (Kaboibus, Wingei, and Jawia) of the East Sepic Province of Papua New Guinea. Results A total of 90 mosquitoes were artificially infected with three strains of Plasmodium : P. vivax- 210 ( n = 30), P. vivax -247 ( n = 30) and P. falciparum ( n = 30). These infected mosquitoes along with another 32 unfed mosquitoes were first checked for the presence of Plasmodium infection by CS-ELISA, and later the same samples were compared with the Cytb -PCR. CS-ELISA for P. vivax -210, P. vivax -247 and P. falciparum detected positive infection in 30, 19 and 18 mosquitoes respectively; whereas Cytb -PCR detected 27, 16 and 16 infections, respectively. The comparison revealed a close agreement between the two assays (κ = 0.862, 0.842 and 0.894, respectively for Pv-210, Pv-247 and P. falciparum groups). It was found ...