PMID: 30141596Dec 1, 2016Paper

Exploration of Using One-step Reverse Transcription PCR in Detection of Four Species of Human Malaria Parasites

Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases
Mei LiZhi-gui Xia

Abstract

To explore the application and specificity of one-step reverse transcription PCR (RT-PCR) in detecting four species of human Plasmodium parasites. Blood samples were collected from a falciparum malaria case, a vivax malaria case, an ovale malaria case, and five quartan malaria cases. RNA and DNA were isolated. One-step RT-PCR and one-step real-time RT-PCR were performed on the RNA digested with DNase to amplify the Plasmodium 18S rRNA. Traditional PCR and one-step RT-PCR were used to amplify 18S rRNAs and 18S rDNAs in differentially diluted RNAs (with or without DNase digestion) and DNAs. The lowest detectable dilution concentrations for the two amplification systems were compared. One-step RT-PCR produced specific bands of 310, 394 and 323 bp, which were sequenced to be 18S rRNA of P. falciparum, P. ovale, and P. vivax. No specific band for P. malariare was found. The one-step real-time RT-PCR results showed fluorescence for all the four species, and all had a melting curve with a single peak except for P. malariare. The lowest detectable dilution concentration by one-step RT-PCR varied from 1 to 10-4 based on the DNA or RNA template amount. Specifically, the lowest detectable dilution concentration of DNA was similar to or l...Continue Reading

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