An Alternative Method for Extracting Plasmodium DNA from EDTA Whole Blood for Malaria Diagnosis

Krongkaew Seesui; Kanokwan Imtawil; Phimphakon Chanetmahun; Porntip Laummaunwai; Thidarut Boonmars

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자료유형: 학술저널

저자정보: Krongkaew Seesui (Khon Kaen University) Kanokwan Imtawil (Khon Kaen University) Phimphakon Chanetmahun (Wiang Haeng Hospital) Porntip Laummaunwai (Khon Kaen University) Thidarut Boonmars (Khon Kaen University)

저널정보: 대한기생충학열대의학회 Parasites, Hosts and Diseases The Korean Journal of Parasitology Vol.56 No.1

발행연도: 2018.2

수록면: 25 - 32 (8page)

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Molecular techniques have been introduced for malaria diagnosis because they offer greater sensitivity and specificity than microscopic examinations. Therefore, DNA isolation methods have been developed for easy preparation and cost effectiveness. The present study described a simple protocol for Plasmodium DNA isolation from EDTA-whole blood. This study demonstrated that after heating infected blood samples with Tris–EDTA buffer and proteinase K solution, without isolation and purification steps, the supernatant can be used as a DNA template for amplification by PCR. The sensitivity of the extracted DNA of Plasmodium falciparum and Plasmodium vivax was separately analyzed by both PCR and semi-nested PCR (Sn-PCR). The results revealed that for PCR the limit of detection was 40 parasites/μl for P. falciparum and 35.2 parasites/μl for P. vivax, whereas for Sn-PCR the limit of detection was 1.6 parasites/μl for P. falciparum and 1.4 parasites/μl for P. vivax. This new method was then verified by DNA extraction of whole blood from 11 asymptomatic Myanmar migrant workers and analyzed by Sn-PCR. The results revealed that DNA can be extracted from all samples, and there were 2 positive samples for Plasmodium (P. falciparum and P. vivax). Therefore, the protocol can be an alternative method for DNA extraction in laboratories with limited resources and a lack of trained technicians for malaria diagnosis. In addition, this protocol can be applied for subclinical cases, and this will be helpful for epidemiology and control.

#Plasmodium falciparum #Plasmodium vivax #EDTA whole blood #malaria diagnosis #DNA extraction #semi-nested PCR

참고문헌 (20)

참고문헌 신청

[보고서] - World Health Organization / 2015 / World Malaria Report, 2015

[학술지(정기간행물)] - Coleman RE / 2006 / Comparison of PCR and microscopy for the detection of asymptomatic malaria in a Plasmodium falciparum/vivax endemic area in Thailand / Malar J 5 : 121 ~

[학술지(정기간행물)] - Ochola LB / 2006 / The reliability of diagnostic techniques in the diagnosis and management of malaria in the absence of a gold standard / Lancet Infect Dis 6 : 582 ~ 588

[학술지(정기간행물)] - Okell LC / 2012 / Factors determining the occurrence of submicroscopic malaria infections and their relevance for control / Nat Commun 3 : 1237 ~

[학술지(정기간행물)] - Rubio JM / 1999 / Semi-nested, multiplex polymerase chain reaction for detection of human malaria parasites and evidence of Plasmodium vivax infection in Equatorial Guinea / Am J Trop Med Hyg 60 : 183 ~ 187

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