Korean J Parasitol.  2016 Jun;54(3):253-259. 10.3347/kjp.2016.54.3.253.

Effective High-Throughput Blood Pooling Strategy before DNA Extraction for Detection of Malaria in Low-Transmission Settings

• Nyunt MH
• Kyaw MP
• Thant KZ
• Shein T
• Han SS
• Zaw NN
• Han JH
• Lee SK
• Muh F
• Kim JY
• Cho SH
• Lee SE
• Yang EJ
• Chang CL
• Han ET

• KMID: 2348265
• DOI: http://doi.org/10.3347/kjp.2016.54.3.253

Abstract

In the era of (pre) elimination setting, the prevalence of malaria has been decreasing in most of the previously endemic areas. Therefore, effective cost- and time-saving validated pooling strategy is needed for detection of malaria in low transmission settings. In this study, optimal pooling numbers and lowest detection limit were assessed using known density samples prepared systematically, followed by genomic DNA extraction and nested PCR. Pooling strategy that composed of 10 samples in 1 pool, 20 µl in 1 sample, was optimal, and the parasite density as low as 2 p/µl for both falciparum and vivax infection was enough for detection of malaria. This pooling method showed effectiveness for handling of a huge number of samples in low transmission settings (<9% positive rate). The results indicated that pooling of the blood samples before DNA extraction followed by usual nested PCR is useful and effective for detection of malaria in screening of hidden cases in low-transmission settings.

Keyword

Plasmodium falciparum; Plasmodium vivax; malaria; pooling strategy; low-transmission setting; nested PCR

MeSH Terms

DNA*
Limit of Detection
Malaria*
Mass Screening
Methods
Parasites
Plasmodium falciparum
Plasmodium vivax
Polymerase Chain Reaction
Prevalence
DNA