A reproducible method for extraction of Plasmodium falciparum DNA by microwave irradiation and its potential for rapid molecular diagnosis
- Author:
Narong Jaturas
;
Thulasi Kumar
;
Brandon Mong Guo Jie
;
Subashini Onichandran
;
Tian Chye Tan
;
Veeranoot Nissapatorn
- Publication Type:Journal Article
- From:Tropical Biomedicine
2015;32(4):753-760
- CountryMalaysia
- Language:English
-
Abstract:
Malaria remains one of the most important communicable diseases. A rapid, simple
and accurate method is a crucial part of malaria diagnosis. The aim of this study was to reevaluate
the microwave irradiation method to extract DNA from Plasmodium falciparum
and compare with six other existing DNA extraction methods such as QIAamp DNA mini kit
(Qiagen), FTA elute card, phenol-chloroform, Chelex, Chelex without proteinase-K and Rapid
boiling. Two different P. falciparum isolates were used: (i) Laboratory strains with 0.3%
parasitemia and (ii) clinical isolate with 0.6% parasitemia. Each DNA extraction method was
validated for the presence of P. falciparum by a routine nested and real time PCR. In order to
evaluate the sensitivity of the DNA extraction by microwave, double serial dilution of P.
falciparum from in vitro culture at parasitemia that ranged from 0.0001 to 0.17% were used
to extract the DNA by microwave and the P. falciparum DNA was then detected by nested
and real-time PCR. The nested and real-time PCR were able to detect. P. falciparum DNA at
the parasitemia level as low as 0.0003% and 0.0001%, respectively. Our results can reproduce
the results from earlier studies and reveal microwave as a rapid and simple tool to extract P.
falciparum DNA and subsequent molecular diagnosis of malaria.
- Full text:P020160316543054417904.pdf
