DNA recovery from archived RDTs for genetic characterization of Plasmodium falciparum in a routine setting in Lambaréné, Gabon
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DNA recovery from archived RDTs for genetic characterization of Plasmodium falciparum in a routine setting in Lambaréné, Gabon. / Nguyen, The Trong; Nzigou Mombo, Brice; Lalremruata, Albert; Koehne, Erik; Zoleko Manego, Rella; Dimessa Mbadinga, Lia Betty; Adegnika, Ayola Akim; Agnandji, Selidji Todagbe; Lell, Bertrand; Kremsner, Peter Gottfried; Velavan, Thirumalaisamy P; Ramharter, Michael; Mordmüller, Benjamin; Mombo-Ngoma, Ghyslain.
In: MALARIA J, Vol. 18, No. 1, 336, 02.10.2019.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - DNA recovery from archived RDTs for genetic characterization of Plasmodium falciparum in a routine setting in Lambaréné, Gabon
AU - Nguyen, The Trong
AU - Nzigou Mombo, Brice
AU - Lalremruata, Albert
AU - Koehne, Erik
AU - Zoleko Manego, Rella
AU - Dimessa Mbadinga, Lia Betty
AU - Adegnika, Ayola Akim
AU - Agnandji, Selidji Todagbe
AU - Lell, Bertrand
AU - Kremsner, Peter Gottfried
AU - Velavan, Thirumalaisamy P
AU - Ramharter, Michael
AU - Mordmüller, Benjamin
AU - Mombo-Ngoma, Ghyslain
PY - 2019/10/2
Y1 - 2019/10/2
N2 - BACKGROUND: Rapid diagnostic tests (RDTs) have been described as a source of genetic material to analyse malaria parasites in proof-of-concept studies. The increasing use of RDTs (e.g., in focal or mass screening and treatment campaigns) makes this approach particularly attractive for large-scale investigations of parasite populations. In this study, the complexity of Plasmodium falciparum infections, parasite load and chloroquine resistance transporter gene mutations were investigated in DNA samples extracted from positive RDTs, obtained in a routine setting and archived at ambient temperature.METHODS: A total of 669 archived RDTs collected from malaria cases in urban, semi-urban and rural areas of central Gabon were used for P. falciparum DNA extraction. Performance of RDTs as a source of DNA for PCR was determined using: (i) amplification of a single copy merozoite surface protein 1 (msp1) gene followed by highly sensitive and automated capillary electrophoresis; (ii) genotyping of the pfcrt gene locus 72-76 using haplotype-specific-probe-based real-time PCR to characterize chloroquine resistance; and, (iii) real-time PCR targeting 18S genes to detect and quantify Plasmodium parasites.RESULTS: Out of the 669 archived RDTs, amplification of P. falciparum nucleic materials had a success rate of 97% for 18S real-time PCR, and 88% for the msp1 gene. The multiplicity of infections (MOI) of the whole population was 2.6 (95% CI 2.5-2.8). The highest number of alleles detected in one infection was 11. The MOI decreased with increasing age (β = - 0.0046, p = 0.02) and residence in Lambaréné was associated with smaller MOIs (p < 0.001). The overall prevalence of mutations associated with chloroquine resistance was 78.5% and was not associated with age. In Lambaréné, prevalence of chloroquine resistance was lower compared to rural Moyen-Ogooué (β = - 0.809, p-value = 0.011).CONCLUSION: RDT is a reliable source of DNA for P. falciparum detection and genotyping assays. Furthermore, the increasing use of RDTs allows them to be an alternative source of DNA for large-scale genetic epidemiological studies. Parasite populations in the study area are highly diverse and prevalence of chloroquine-resistant P. falciparum remains high, especially in rural areas.
AB - BACKGROUND: Rapid diagnostic tests (RDTs) have been described as a source of genetic material to analyse malaria parasites in proof-of-concept studies. The increasing use of RDTs (e.g., in focal or mass screening and treatment campaigns) makes this approach particularly attractive for large-scale investigations of parasite populations. In this study, the complexity of Plasmodium falciparum infections, parasite load and chloroquine resistance transporter gene mutations were investigated in DNA samples extracted from positive RDTs, obtained in a routine setting and archived at ambient temperature.METHODS: A total of 669 archived RDTs collected from malaria cases in urban, semi-urban and rural areas of central Gabon were used for P. falciparum DNA extraction. Performance of RDTs as a source of DNA for PCR was determined using: (i) amplification of a single copy merozoite surface protein 1 (msp1) gene followed by highly sensitive and automated capillary electrophoresis; (ii) genotyping of the pfcrt gene locus 72-76 using haplotype-specific-probe-based real-time PCR to characterize chloroquine resistance; and, (iii) real-time PCR targeting 18S genes to detect and quantify Plasmodium parasites.RESULTS: Out of the 669 archived RDTs, amplification of P. falciparum nucleic materials had a success rate of 97% for 18S real-time PCR, and 88% for the msp1 gene. The multiplicity of infections (MOI) of the whole population was 2.6 (95% CI 2.5-2.8). The highest number of alleles detected in one infection was 11. The MOI decreased with increasing age (β = - 0.0046, p = 0.02) and residence in Lambaréné was associated with smaller MOIs (p < 0.001). The overall prevalence of mutations associated with chloroquine resistance was 78.5% and was not associated with age. In Lambaréné, prevalence of chloroquine resistance was lower compared to rural Moyen-Ogooué (β = - 0.809, p-value = 0.011).CONCLUSION: RDT is a reliable source of DNA for P. falciparum detection and genotyping assays. Furthermore, the increasing use of RDTs allows them to be an alternative source of DNA for large-scale genetic epidemiological studies. Parasite populations in the study area are highly diverse and prevalence of chloroquine-resistant P. falciparum remains high, especially in rural areas.
KW - Adolescent
KW - Adult
KW - Biological Specimen Banks
KW - Body Temperature
KW - Child
KW - Child, Preschool
KW - Chloroquine/pharmacology
KW - DNA, Protozoan/genetics
KW - Drug Resistance/genetics
KW - Female
KW - Gabon
KW - Genotype
KW - Humans
KW - Malaria, Falciparum/blood
KW - Male
KW - Membrane Transport Proteins/genetics
KW - Merozoite Surface Protein 1/genetics
KW - Molecular Diagnostic Techniques
KW - Parasitemia
KW - Plasmodium falciparum/drug effects
KW - Protozoan Proteins/genetics
KW - Retrospective Studies
KW - Young Adult
U2 - 10.1186/s12936-019-2972-y
DO - 10.1186/s12936-019-2972-y
M3 - SCORING: Journal article
C2 - 31578142
VL - 18
JO - MALARIA J
JF - MALARIA J
SN - 1475-2875
IS - 1
M1 - 336
ER -