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Prevalence of pfdhfr and pfdhps mutations in Plasmodium falciparum associated with drug resistance among pregnant women receiving IPTp-SP at Msambweni County Referral Hospital, Kwale County, Kenya.

ABSTRACT: BACKGROUND:Prevention and treatment of malaria during pregnancy is crucial in dealing with maternal mortality and adverse fetal outcomes. The World Health Organization recommendation to treat all pregnant women with sulfadoxine-pyrimethamine (SP) through antenatal care structures was implemented in Kenya in the year 1998, but concerns about its effectiveness in preventing malaria in pregnancy has arisen due to the spread of SP resistant parasites. This study aimed to determine the prevalence of SP resistance markers in Plasmodium falciparum parasites isolated from pregnant women seeking antenatal care at Msambweni County Referral Hospital, located in coastal Kenya, between the year 2013 and 2015. METHODS:This hospital-based study included 106 malaria positive whole blood samples for analysis of SP resistance markers within the Pfdhfr gene (codons 51, 59 and 108) and Pfdhps gene (codons 437 and 540). The venous blood collected from all pregnant women was tested for malaria via light microscopy, then the malaria positive samples were separated into plasma and red cells and stored in a - 86° freezer for further studies. Archived red blood cells were processed for molecular characterization of SP resistance markers within the Pfdhfr and Pfdhps genes using real time PCR platform and Sanger sequencing. RESULTS:All samples had at least one mutation in the genes associated with drug resistance; polymorphism prevalence of Pfdhfr51I, 59R and 108N was at 88.7%, 78.3% and 93.4%, respectively, while Pfdhps polymorphism accounted for 94.3% and 91.5% at 437G and 540E, respectively. Quintuple mutations (at all the five codons) conferring total SP resistance had the highest prevalence of 85.8%. Quadruple mutations were observed at a frequency of 10.4%, and 24.5% had a mixed outcome of both wildtype and mutant genotypes in the genes of interest. CONCLUSION:The data suggest a high prevalence of P. falciparum genetic variations conferring resistance to SP among pregnant women, which may explain reduced efficacy of IPTp treatment in Kenya. There is need for extensive SP resistance profiling in Kenya to inform IPTp drug choices for successful malaria prevention during pregnancy.

SUBMITTER: Gikunju SW

PROVIDER: S-EPMC7247205 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Prevalence of pfdhfr and pfdhps mutations in Plasmodium falciparum associated with drug resistance among pregnant women receiving IPTp-SP at Msambweni County Referral Hospital, Kwale County, Kenya.

Gikunju Stella Wanjiku SW Agola Eric L EL Ondondo Raphael Omusebe RO Kinyua Johnson J Kimani Francis F LaBeaud Angelle Desiree AD Malhotra Indu I King Charles C Thiong'o Kelvin K Mutuku Francis F

Malaria journal 20200524 1

<h4>Background</h4>Prevention and treatment of malaria during pregnancy is crucial in dealing with maternal mortality and adverse fetal outcomes. The World Health Organization recommendation to treat all pregnant women with sulfadoxine-pyrimethamine (SP) through antenatal care structures was implemented in Kenya in the year 1998, but concerns about its effectiveness in preventing malaria in pregnancy has arisen due to the spread of SP resistant parasites. This study aimed to determine the preval ...[more]

PMID: 32448228

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Project description:BackgroundArbovirus surveillance and recurrence of outbreaks in Kenya continues to reveal the re-emergence of viruses of public health importance. This calls for sustained efforts in early detection and characterization of these agents to avert future potential outbreaks.MethodsA larval survey was carried out in three different sites in Kwale County, Vanga, Jego and Lunga Lunga. All containers in every accessible household and compound were sampled for immature mosquitoes. In addition, adult mosquitoes were also sampled using CO2-baited CDC light traps and BG-Sentinel traps in the three sites and also in Tsuini. The mosquitoes were knocked down using trimethylamine and stored in a liquid nitrogen shipper for transportation to the laboratory where they were identified to species, pooled and homogenized ready for testing.ResultsA total of 366 houses and 1730 containers were inspected. The House Index (HI), Container Index (CI) and Breateau Index (BI) for Vanga Island were (3%: 0.66: 3.66) respectively. In Jego, a rural site, the HI, CI and BI were (2.4%: 0.48: 2.4) respectively. In Lunga Lunga, a site in an urban area, the HI, CI and BI were (22.03%: 3.97: 29.7) respectively. The indices suggest that this region is at risk of arbovirus transmission given they were above the WHO threshold (CI > 1, HI > 1% and BI > 5). The most productive containers were the concrete tanks (44.4%), plastic tank (22.2%), claypot (13.3%), plastic drums (8.9%), plastic basins (4%), jerricans (1.2%) and buckets (0.3%). Over 20,200 adult mosquitoes were collected using CDC light traps, and over 9,200 using BG- sentinel traps. These mosquitoes were screened for viruses by inoculating in Vero cells. Eleven Orthobunyavirus isolates were obtained from pools of Ae. pembaensis (4), Ae. tricholabis (1), Cx. quinquefasciatus (3), Culex spp. (1) and Cx. zombaensis (2). Five of the Orthobunyaviruses were sequenced and four of these were determined to be Bunyamwera viruses while one isolate was found to be Nyando virus. One isolate remained unidentified.ConclusionsThese results indicate circulation of Orthobunyaviruses known to cause diverse grades of febrile illness with rash in humans in this region and highlights the need for continued monitoring and surveillance to avert outbreaks.

Project description:Despite the extensive ownership and use of insecticide-treated nets (ITNs) over the last decade, the effective lifespan of these nets, especially their physical integrity, under true operational conditions is not well-understood. Usefulness of nets declines primarily due to physical damage or loss of insecticidal activity.A community based cross-sectional survey was used to determine the physical condition and to identify predictors of poor physical condition for bed nets owned by individuals from communities in Kwale County, coastal Kenya. A proportionate hole index (pHI) was used as a standard measure, and the cut-offs for an 'effective net' (offer substantial protection against mosquito bites) and 'ineffective nets' (offer little or no protection against mosquito bites) were determined (pHI ?88 (about ?500 cm2 of holes surface area) and pHI of >88 (?500 cm2 of holes surface area), respectively).The vast majority (78%) of the surveyed nets had some holes. The median pHI was 92 (range: 1-2,980). Overall, half of the nets were categorized as 'effective nets' or 'serviceable nets'. Physical deterioration of nets was associated with higher use and washing frequency. Young children and older children were found to use ineffective bed nets significantly more often than infants, while the physical integrity of nets owned by pregnant women was similar to those owned by infants. Estuarine environment inhabitants owned nets with the worst physical condition, while nets owned by the coastal slope inhabitants were in fairly good physical condition. The results suggest that bed nets are optimally utilized when they are new and physically intact. Thereafter, bed net utilization decreases gradually with increasing physical deterioration, with most net owners withdrawing physically damaged nets from routine use.This withdrawal commonly happens following 1.5 years of use, making bed net use the most important predictor of physical integrity. On average, the nets were washed twice within six months prior to the survey. Washing frequency was significantly influenced by the bed net colour and bed net age. Lack of knowledge on reasons for net retreatment and the retreatment procedure was evident, while net repair was minimal and did not seem to improve the physical condition of the nets. The "catch-up" bed net distribution strategies are sufficient for ensuring adequate ownership and utilization of 'effective nets' in the targeted groups, but bi-annual mass distribution is necessary to provide similar ownership and utilization for the other groups not targeted by "catch-up" strategies.Monitoring and maintenance strategies that will deliver locally appropriate education messages on net washing and repair will enhance the effectiveness of malaria control programmes, and further research to assess ineffective nets need is needed.

Project description:BackgroundBoth pre-donation and post-donation deferrals pose challenges to blood safety and availability. This study delved into the deferral rates before donations and their underlying reasons, as, transfusion transmissible infections (TTIs) leading to post-donation deferrals among potential blood donors at the Kwale Satellite Blood Transfusion Centre (KSBTC) in Kenya.MethodsWe performed a retrospective electronic record review of pre- and post-donation deferrals among blood donors at KSBTC, 2018-2022. The pre-donations deferral rate and reasons for deferral were analyzed. Accepted donations were analyzed to determine the prevalence of HIV, hepatitis B (HBV), hepatitis C (HCV), and syphilis. Descriptive statistics were calculated and both crude odds ratio (COR) and adjusted odds ratio (AOR), and their 95% confidence intervals (CI) were calculated. Variables with p < 0.05 were considered statistically significant.ResultsA review was conducted on 12,633 blood donation records. Among these, individuals 2,729/12,633 (21.60%) were deferred from donating with the primary reason being low hemoglobin levels, constituting 51.86% of deferrals. Around 773/9,904 (7.80%) of blood units, were discarded due to at least one TTI. Among these, HBV accounted for 4.73%, HIV for 2.01%, HCV for 1.21%, and Syphilis for 0.59% of cases. The adjusted odds ratio for male donors were, (aOR = 1.3, 95% CI 1.01-1.57), donors with none or primary education level (aOR = 1.4 95% CI 1.11-1.68), first-timer donors (aOR = 1.2, 95% CI 1.01-1.44), and static strategy for blood collection (aOR = 1.4, 95%CI 1.12-1.63) were independently potentially associated with testing positive for at least one TTI.ConclusionThe study indicates that TTIs continue to pose a risk to the safety of Kenya's bloodstock, with a notable prevalence of HBV infections. Male donors, individuals with limited education, first-time donors, and utilizing a fixed strategy for blood collection were identified as potential risk factors independently associated with TTIs.

Project description:BackgroundMyths and misconceptions around modern contraceptives have been associated with low contraceptive uptake in sub-Saharan Africa and Kenya in particular. Addressing persistent contraceptive knowledge gaps can make a significant contribution towards improved contraceptive uptake among young women. This qualitative study therefore sought to explore and understand young people's knowledge of modern contraception and to identify their key concerns regarding these methods.MethodsWe used focus group discussions (FGD) with vignette and writing activities to explore key myths and misconceptions around the use of contraceptives. Six FGDs (three for young men and three for young women) were conducted with a total of 28 young women and 30 young men from Kwale County, Kenya. We included 10 discussants aged 18-24 per FGD, one FGD had 8 participants. Predefined codes reflecting the discussion guides and emerging issues in the FGDs were used to develop the thematic coding framework. Our analysis followed a pattern of association on the key preset themes focusing on myths and misconceptions around contraceptive use.ResultsResults are presented under four key themes: awareness of contraception, myths and misconceptions around contraception, males' contraceptive narratives and young people's preferred sources of contraceptives. Both men and women participants reported basic awareness of contraceptives. A mixture of biological and social misconceptions were discussed and included perceptions that modern contraception: jeopardized future fertility, could result in problems conceiving or birth defects, made women promiscuous, was 'un-African', and would deny couples their sexual freedom. Compared to female respondents in the study, young men appeared to be strong believers of the perceived socio-cultural effects of contraceptives. On preferred sources of contraceptives, respondents reported on two main sources, pharmacies and public hospitals, however, they could not agree on which one was suitable for them.ConclusionsThis study revealed the presence of a mixture of biological and social myths and misconceptions around contraception, with young men also strongly adhering to these misconceptions. The low level of contraceptive knowledge, particularly on contraceptive fears as revealed by the study demonstrate critical gaps in sexual and reproductive health (SRH) knowledge among young people. Improved SRH literacy to address contraceptives' fears through appropriate and gender specific interventions to reach out to young men and women with factual SRH information may therefore contribute to increased uptake of SRH services including modern contraceptive methods.

Project description:BackgroundThe amplification of GTP cyclohydrolase 1 (pfgch1) in Plasmodium falciparum has been linked to the upregulation of the pfdhfr and pfdhps genes associated with resistance to the antimalarial drug sulfadoxine-pyrimethamine. During the 1990s and 2000s, sulfadoxine-pyrimethamine was withdrawn from use as first-line treatment in southeast Asia due to clinical drug resistance. This study assessed the temporal and geographic changes in the prevalence of pfdhfr and pfdhps gene mutations and pfgch1 amplification a decade after sulfadoxine-pyrimethamine had no longer been widely used.MethodsA total of 536 P. falciparum isolates collected from clinical trials in Thailand, Cambodia, and Lao PDR between 2008 and 2018 were assayed. Single nucleotide polymorphisms of the pfdhfr and pfdhps genes were analyzed using nested PCR and Sanger sequencing. Gene copy number variations of pfgch1 were investigated using real-time polymerase chain reaction assay.ResultsSequences of the pfdhfr and pfdhps genes were obtained from 96% (517/536) and 91% (486/536) of the samples, respectively. There were 59 distinct haplotypes, including single to octuple mutations. The two major haplotypes observed included IRNI-AGEAA (25%) and IRNL-SGKGA (19%). The sextuple mutation IRNL-SGKGA increased markedly over time in several study sites, including Pailin, Preah Vihear, Ratanakiri, and Ubon Ratchathani, whereas IRNI-AGEAA decreased over time in Preah Vihear, Champasak, and Ubon Ratchathani. Octuple mutations were first observed in west Cambodia in 2011 and subsequently in northeast Cambodia, as well as in southern Laos by 2018. Amplification of the pfgch1 gene increased over time across the region, particularly in northeast Thailand close to the border with Laos and Cambodia.ConclusionDespite the fact that SP therapy was discontinued in Thailand, Cambodia, and Laos decades ago, parasites retained the pfdhfr and pfdhps mutations. Numerous haplotypes were found to be prevalent among the parasites. Frequent monitoring of pfdhfr and pfdhps in these areas is required due to the relatively rapid evolution of mutation patterns.

Project description:BackgroundAnti-malarial drug resistance in Kenya prompted two drug policy changes within a decade: sulphadoxine-pyrimethamine (SP) replaced chloroquine (CQ) as the first-line anti-malarial in 1998 and artemether-lumefantrine (AL) replaced SP in 2004. Two cross-sectional studies were conducted to monitor changes in the prevalence of molecular markers of drug resistance over the period in which SP was used as the first-line anti-malarial. The baseline study was carried out from 1999-2000, shortly after implementation of SP, and the follow-up study occurred from 2003-2005, during the transition to AL.Materials and methodsBlood was collected from malaria smear-positive, symptomatic patients presenting to outpatient centers in Kisumu, Kenya, during the baseline and follow-up studies. Isolates were genotyped at codons associated with SP and CQ resistance. In vitro IC50 values for antifolates and quinolones were determined for isolates from the follow-up study.ResultsThe prevalence of isolates containing the pfdhfr N51I/C59R/S108N/pfdhps A437G/K540E quintuple mutant associated with SP-resistance rose from 21% in the baseline study to 53% in the follow-up study (p < 0.001). Isolates containing the pfdhfr I164L mutation were absent from both studies. The pfdhps mutations A581G and A613S/T were absent from the baseline study but were present in 85% and 61%, respectively, of isolates from the follow-up study. At follow-up, parasites with mutations at five pfdhps codons, 436, 437, 540, 581, and 613, accounted for 39% of isolates. The CQ resistance-associated mutations pfcrt K76T and pfmdr1 N86Y rose from 82% to 97% (p = 0.001) and 44% to 76% (p < 0.001), respectively, from baseline to follow-up.ConclusionsDuring the period in which SP was the first-line anti-malarial in Kenya, highly SP-resistant parasites emerged, including isolates harboring pfdhps mutations not previously observed there. SP continues to be widely used in Kenya; however, given the highly resistant genotypes observed in this study, its use as a first-line anti-malarial should be discouraged, particularly for populations without acquired immunity to malaria. The increase in the pfcrt K76T prevalence, despite efforts to reduce CQ use, suggests that either these efforts are not adequate to alleviate CQ pressure in Kisumu, or that drug pressure is derived from another source, such as the second-line anti-malarial amodiaquine.

Dataset Information

Prevalence of pfdhfr and pfdhps mutations in Plasmodium falciparum associated with drug resistance among pregnant women receiving IPTp-SP at Msambweni County Referral Hospital, Kwale County, Kenya.

Publications

Prevalence of pfdhfr and pfdhps mutations in Plasmodium falciparum associated with drug resistance among pregnant women receiving IPTp-SP at Msambweni County Referral Hospital, Kwale County, Kenya.

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