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. 2016 Jun 27;9(1):362.
doi: 10.1186/s13071-016-1641-z.

Multiplicity of infection and genetic diversity of Plasmodium falciparum isolates from patients with uncomplicated and severe malaria in Gezira State, Sudan

Muzamil Mahdi Abdel Hamid  1 , Arwa F Elamin  2 , Musab M Ali Albsheer  2 , Abdelmohaymin A A Abdalla  3 , Nouh S Mahgoub  2 , Shaza O Mustafa  2 , Mohamed SiddigEltayeb Muneer  3 , Mutaz Amin  3
Affiliations

Multiplicity of infection and genetic diversity of Plasmodium falciparum isolates from patients with uncomplicated and severe malaria in Gezira State, Sudan

Muzamil Mahdi Abdel Hamid et al. Parasit Vectors. .

Abstract

Background: Multiplicity and genetic diversity of Plasmodium falciparum infection might play a role in determining the clinical outcome of malaria infection and could be a fair reflection of the disease transmission rate. This study investigated the genetic diversity of P. falciparum and multiplicity of infection in relation to the severity of malaria and age of patients in Gezira State, Sudan.

Methods: A cross-sectional health facilities-based survey was conducted in Gezira State, Sudan in January 2012. A total of 140 P. falciparum malaria patients diagnosed with microscopy and confirmed using nested PCR were recruited and classified into uncomplicated malaria and severe malaria states according to the standard WHO criteria. DNA was extracted and MSP1 and MSP2 allelic families were determined using nested PCR.

Results: The overall multiplicity of infection (MOI) was 2.25 and 2.30 and 2.15 for uncomplicated and severe malaria respectively. There were no statistically significant differences between uncomplicated and severe malaria (SM) patient groups in MOI with regard to MSP1, MSP2 and overall MOI (Mann-Whitney U-test; all P < 0.05). The predominant MSP1 allelic families were MAD20 for uncomplicated malaria and RO33 for severe malaria. The distribution of both FC27 and IC1/3D7 MSP2 allelic families were approximately the same across disease severity. One hundred and eleven P. falciparum isolates (81 %) consisted of multiple genotypes; 71/90 (78.9 %) in uncomplicated malaria and 40/50 (85.1 %) in severe malaria patient groups. Neither MSP1 nor MSP2 allelic families showed association with malaria severity. No statistically significant differences in multiplicity of infection were observed between different age groups.

Conclusion: In this study the majority of P. falciparum isolates from uncomplicated and severe malaria patients consisted of multiple genotypes. Further molecular epidemiological studies delineate the link between P. falciparum genotype with the malaria phenotype in different regions are encouraged.

Keywords: Genetic diversity; MSP1; MSP2; Multiplicity of infection; Plasmodium falciparum; Severity; Sudan.

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Figures

Fig. 1
Fig. 1
Gel electrophoresis picture for detection of 18S rRNA P. falciparum gene: Lane 1, 100 bp DNA ladder; Lanes 2–5, 200 bp18S rRNA gene; Lane 6, negative control
Fig. 2
Fig. 2
Gel electrophoresis of MSP1 allelic types: Lane 1,100 bp DNA ladder; l. Lanes 2 and 3, 100 bp and 180 bp of MAD20; Lanes 4 and 5, 100 bp and 220 bp of MAD20; Lane 6, negative control of MAD20; Lanes 7 and 8, negative samples of K1; Lanes 9 and 10, 180 bp and 200 bp of K1; Lane 11, negative control of K1; Lanes 12–14, 160 bp of RO33; Lane 15, negative control of RO33
Fig. 3
Fig. 3
Gel electrophoresis of MSP2 allelic types: Lane 1, 100 bp DNA ladder; Lanes 2–4, 300 bp and 350 bp of FC27; Lane 5, 500 bp and 550 bp and of IC1/3D7; Lane 6, 500 bp of ICI/3D7; Lane 7, negative control
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