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PHD : Parasites, Hosts and Diseases

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Original Article

Imported Malaria in United Arab Emirates: Evaluation of a New DNA Extraction Technique Using Nested PCR

The Korean Journal of Parasitology 2009;47(3):227-233.
Published online: August 28, 2009

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Corresponding author (doaa68@yahoo.com)
• Received: February 11, 2009   • Revised: April 29, 2009   • Accepted: May 16, 2009

Copyright © 2009 by The Korean Society for Parasitology

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Imported Malaria in United Arab Emirates: Evaluation of a New DNA Extraction Technique Using Nested PCR
Korean J Parasitol. 2009;47(3):227-233.   Published online August 28, 2009
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Imported Malaria in United Arab Emirates: Evaluation of a New DNA Extraction Technique Using Nested PCR
Korean J Parasitol. 2009;47(3):227-233.   Published online August 28, 2009
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Imported Malaria in United Arab Emirates: Evaluation of a New DNA Extraction Technique Using Nested PCR
Image Image
Fig. 1 Comparative sensitivity of different DNA extraction techniques for Plasmodium genus detection.
Fig. 2 DNA extracted from malaria patients and amplified by nested PCR. Assays of sensitivities, using genus- and species-specific primer sets. (A) DNA by FTA Elute card. Lanes 1 & 3-11, positive samples, Plasmodium genus fragment (240 bp); 2, molecular marker 100 bp (Promega). (B) DNA by rapid boiling technique. Lanes 2-6 & 13-14, Plasmodium genus fragment (240 bp). DNA by FTA classic cards. Lanes 7-12, Plasmodium genus fragment (240 bp). Lane 1, molecular marker 100 bp (Promega). (C) Species-specific primers. Lanes 2-4, P. vivax fragment (117 bp); 5-15, negative results; 1, molecular marker 100 bp (Promega). (D) Species-specific primers in 3 samples; sample 1, lanes 1-4; sample 2, lanes 10-13 showing mixed infections with P. falciparum and P. vivax; sample 3, lanes 6-9 showing single infection with P. falciparum. Each sample was run against the 4 species primer sets. Lanes 1, P. falciparum fragment (205 bp); 2, P. ovale primer (negative); 3, P. malariae primer (negative); 4, P. vivax fragment (117 bp); 5, molecular marker 100 bp (Promega); 6, P. falciparum fragment (205 bp); 7, P. ovale primer (negative); 8, P. malariae primer (negative); 9, P. vivax fragment (negative); 10, P. falciparum fragment (205 bp); 11, P. ovale primer (negative); 12, P. malariae primer (negative); 13, P. vivax fragment (117 bp).
Imported Malaria in United Arab Emirates: Evaluation of a New DNA Extraction Technique Using Nested PCR
Results No. positive (%) No. negative (%) Sensitivity (%)* Microscopy 52 (39.4) 80 (60.6) 88.2 ELISA 46 (34.8) 86 (65.2) 81.1 PCR 60 (45.5) 72 (54.5) 100.0 Microscopy Nested PCR
Nested PCR total Negative Pv Pf Negative 72 0 0 72 Pv 4 31 0 35 Pf 4 0 17 21 Pv + Pf 0 2 2 4 Microscopy total 80 33 19 132 Number of cases PCR result Microscopy result Repeated PCR result Repeated microscopy result 1 Pv Negative Pv Pv 2 Pv Negative Pv Pv 3 Pf Negative Pf Pf 4 Pv Negative Pv Negative 5 Pv + Pf Pf Pv + Pf Pf 6 Pf Negative Pf Negative 7 Pv Negative Pv Pv 8 Pv + Pf Pv Pv + Pf Pv 9 Pf Negative Pf Pf 10 Pv + Pf Pf Pv + Pf Pv 11 Pf Negative Pf Pf 12 Pv + Pf Pv Pv + Pf Pv
Table 1. Sensitivity of microscopy, ELISA, and PCR methods in the Plasmodium genus detection

PCR was used as the gold standard.

Table 2. Comparison of the genus and species detection using microscopy and PCR

Pv, Plasmodium vivax; Pf, Plasmodium falciparum; Pv + Pf, Mixed infection with P. vivax and P. falciparum.

Table 3. Discordant results using microscopy and PCR

Pv, Plasmodium vivax; Pf, Plasmodium falciparum; Pv + Pf, Mixed infection with P. vivax and P. falciparum.