Detection of Genetic markers associated with Plasmodium falciparum drug resistance among malaria patients in Kaduna State, Nigeria

2021 ◽  
Vol 42 (2) ◽  
pp. 206-213
Author(s):  
G.Y. Benjamin ◽  
H.I. Inabo ◽  
M.H.I. Doko ◽  
B.O. Olayinka

Malaria is a disease of public health concern in Nigeria and sub-Saharan Africa. It is caused by intracellular parasites of the genus Plasmodium. The aim of this study was to detect genetic markers associated with Plasmodium falciparum drug resistance among malaria patients in Kaduna State, Nigeria. The study was a cross-sectional study that lasted from May 2018 to October 2018. Three hundred blood samples were collected from consenting individuals attending selected hospitals, in the three senatorial districts of Kaduna State, Nigeria. Structured questionnaire were used to obtain relevant data from study participants. The blood samples were screened for malaria parasites using microscopy and rapid diagnostic test kit. Polymerase Chain Reaction was used for detection of the drug resistance genes. Pfcrt, pfmdr1, pfdhfr, pfdhps and pfatpase6 genes were detected at expected amplicon sizes from the malaria positive samples. The pfatpase6 PCR amplicons were sequenced and a phylogenetic tree was created to determine their relatedness. Result showed that Pfcrt (80%) had the highest prevalence, followed by pfdhfr (60%), pfmdr1 (36%) and pfdhps (8%). Pfatpase6 was also detected in 73.3% of the samples, and a phylogenetic tree showed relatedness between the pfatpase6  sequences in this study and those deposited in the GenBank. In conclusion, the study detected that Plasmodium falciparum genes were associated with drug resistance to commonly used antimalarials.

2020 ◽  
Author(s):  
Gideon Yakusak Benjamin ◽  
Helen Ileigo Inabo ◽  
Hassan Isa Doko Muhammad ◽  
Busayo O Olayinka

Abstract Background Malaria is a disease of public health concern in Nigeria and sub-Saharan Africa. The emergence of drug resistance, particularly among P. falciparum strains, has been a major contributor to the global burden of malaria. This research was aimed at detecting genetic markers (pfcrt, pfmdr1, pfdhfr, pfdhps, pfatpase6) associated with antimalarial drug resistance and assessing the distribution of haemoglobin genotypes among malaria patients in of Kaduna State, Nigeria. Methods Three hundred (300) blood samples were collected from consenting individuals attending selected hospitals, in the three senatorial districts of Kaduna State, Nigeria. A structured questionnaire was used to obtain relevant data from the study participants. The samples were screened for malaria parasites by microscopy and malaria rapid diagnostic test kit. Deoxyribonucleic acid was extracted from one third of the malaria positive samples, and Polymerase Chain Reaction (PCR) was used for detection of the drug resistance genes. Pfcrt, pfmdr1, pfdhfr, pfdhps and pfatpase6 genes were detected at expected amplicon sizes from the malaria positive samples. The pfatpase6 PCR amplicons were sequenced and a phylogenetic tree was created using MEGA X to determine their relatedness to published sequences. Results Pfcrt (80%) had the highest prevalence, followed by pfdhfr (60%), pfmdr1 (36%) and pfdhps (8%). Pfatpase6 was also detected in 73.3% of the samples. The phylogenetic tree showed that all the pfatpase6 gene sequences (both the ones from this study and those published in NCBI Genbank) had the same origin and were closely related. However, the sequences from NCBI Genbank were from one clade; arising from a common ancestor (monophyletic) thus they were more closely related to themselves, than to the pfatpase6 sequences obtained in this study. Of all the malaria positive participants, those with HbAA (73%) haemoglobin genotype had the highest percentage followed by HbAS (23%), HbAC (3%) and HbSS (1). Conclusion We detected Plasmodium falciparum genes associated with drug resistance to commonly used antimalarials in the study area. Expression of these genes could have serious consequences in the treatment of malaria. The percentage of Plasmodium falciparum malaria was higher among persons with HbAA than those with HbAS, HbAC and HbS.


2020 ◽  
Author(s):  
Gideon Yakusak Benjamin ◽  
Helen Ileigo Inabo ◽  
Hassan Isa Doko Muhammad ◽  
Busayo O Olayinka

Abstract Background Malaria is a disease of public health concern in Nigeria and sub-Saharan Africa. The emergence of drug resistance, particularly among P. falciparum strains, has been a major contributor to the global burden of malaria. This research was aimed at detecting genetic markers associated with antimalarial drug resistance and assessing the distribution of haemoglobin genotypes among malaria patients in of Kaduna State, Nigeria.Methods Three hundred (300) blood samples were collected from consenting individuals attending selected hospitals, in the three senatorial districts of Kaduna State, Nigeria. A structured questionnaire was used to obtain relevant data from the study participants. The samples were screened for malaria parasites by microscopy and malaria rapid diagnostic test kit. Deoxyribonucleic acid was extracted from one third of the malaria positive samples, and Polymerase Chain Reaction (PCR) was used for detection of the drug resistance genes. Pfcrt, pfmdr1, pfdhfr, pfdhps and pfatpase6 genes were detected at expected amplicon sizes from the malaria positive samples. The pfatpase6 PCR amplicons were sequenced and a phylogenetic tree was created using MEGA X to determine their relatedness to published sequences.Results Pfcrt (80%) had the highest prevalence, followed by pfdhfr (60%), pfmdr1 (36%) and pfdhps (8%). Pfatpase6 was also detected in 73.3% of the samples. The phylogenetic tree showed that all the pfatpase6 gene sequences (both the ones from this study and those published in NCBI Genbank) had the same origin and were closely related. However, the sequences from NCBI Genbank were from one clade; arising from a common ancestor (monophyletic) thus they were more closely related to themselves, than to the pfatpase6 sequences obtained in this study. Of all the malaria positive participants, those with HbAA (73%) haemoglobin genotype had the highest percentage followed by HbAS (23%), HbAC (3%) and HbSS (1).Conclusion We detected Plasmodium falciparum genes associated with drug resistance to commonly used antimalarials in the study area. Expression of these genes could have serious consequences in the treatment of malaria. Persons with HbAS, HbAC and HbSS may enjoy some protection from falciparum malaria than those with HbAA.


2019 ◽  
Vol 4 (Suppl 3) ◽  
pp. A53.3-A53
Author(s):  
Rella Zoleko Manego ◽  
Eric Koehne ◽  
Ghyslain Mombo-Ngoma

BackgroundMalaria remains a public health issue particularly in sub-Saharan Africa with special features of seriousness in young children and pregnant women. Older children and adults reported to have acquired a semi-immune status and therefore show low parasitaemia and less pronounced symptoms if any. We aim to explore parasitaemia and symptoms in adolescents and adults with malaria compared to younger children in the Lambaréné area.MethodsWe conducted a cross-sectional study in CERMEL for a month in 2018. Adults and children were screened for malaria at health facilities in Lambaréné and Fougamou and their respective surrounding villages in the central region of Gabon. Malaria infection was determined either by rapid diagnostic test (RDT) or by thick blood smear (TBS). Body temperature, history of fever, age, sex, haemoglobin level, and location were collected.ResultsFrom 09 January to 09 February 2018, a total of 596 patients were screened and 280 (47%) belonged to the adult population. Plasmodium falciparum infection in adults was diagnosed based on the detection of P. falciparum in thick blood films. Prevalence among the adults was 56% (n=141); 34% of adult patients had a parasite count between 1015–116,079 pf/µl. Fever was found in 31% of adults and several symptoms were reported by the same patient. Vomiting, loss of appetite, and fatigue were significantly more present in the adult population than in children.ConclusionThis study shows a high rate of Plasmodium falciparum infection in adults associated to clinical symptoms and high level of parasitaemia. Adults seem to become an at-risk population, suggesting that malaria programmes should consider including adults in the implementation of a malaria prevention programme.


2019 ◽  
Vol 4 (Suppl 3) ◽  
pp. A40.1-A40
Author(s):  
Dossou Akpéyédjé ◽  
Saadou Issifou

BackgroundIn sub-Saharan Africa, malaria during pregnancy is a major health problem because it poses significant risks for the pregnant woman and the foetus. The sequestration of Plasmodium falciparum-infected erythrocytes in the placenta has consequences for the mother and the foetus. This study aimed to evaluate the allelic polymorphism of the Plasmodium falciparum MSP-2 gene related to the consequences of placental malaria.MethodsIt was a cross-sectional study conducted over two periods lasting six months in 2016 and 2017. The maternity center of the Hospital of Borgou-Alibori in Benin served as a framework for the study. From the 98 parturients included, placental blood samples were taken and then genotyped.ResultsUsing the MSP-2 gene as marker, the prevalence was 17, 34%. The MSP-2 gene was polymorphic with 9 distinct allelic types for both 3D7 and FC27 families (150 bp; 200 bp; 250 bp; 275 bp; 300 bp; 350 bp; 400 bp; 450 bp and 500 bp). The FC27 allelic family was predominant over the 3D7 family with 56, 25% and 43, 75% respectively. The 300 bp allelic type (50%) was predominant in the FC27 family while the 400 bp type was predominant in 3D7 family (35, 71%). 9 women had polyclonality (52,94%). The multiplicity of infection (MOI) was 1, 88. The number of strains ranged 1 to 4 in infected women. In univariate analysis there was no significant relationship between MSP-2 gene polymorphism and maternofoetal consequences. The absence of „prenatal consultation (p=0.0270), non-taking of IPTp/SP (p=0.0060), the occurrence of malaria in the third trimester (p=0,0364) and moderate maternal anaemia (p=0.0277) were associated with the polymorphism of MSP-2 gene. The MOI was significantly associated with parasite density of infected women.ConclusionPlasmodium falciparum MSP-2 gene was polymorphic in infected women at Parakou. Several factors related to pregnancy monitoring were associated with this genetic diversity. It is therefore essential to ensure correct follow-up of pregnancies.


2019 ◽  
Vol 2019 ◽  
pp. 1-8
Author(s):  
Jeanne Vanessa Koumba Lengongo ◽  
Yaye Dié Ndiaye ◽  
Marie Louise Tshibola Mbuyi ◽  
Jacques Mari Ndong Ngomo ◽  
Daouda Ndiaye ◽  
...  

Background. Studying malaria parasites cross resistance to sulfadoxine-pyrimethamine (SP) and trimethoprim-sulfamethoxazole (cotrimoxazole, CTX) is necessary in areas coendemic for malaria and HIV. Polymorphism and frequency of drug resistance molecular markers, Pfdhfr and Pfdhps genes have been assessed in Plasmodium falciparum isolates from HIV-infected adults, in Gabon. Materiel and Methods. A cross-sectional study was conducted in three HIV care and treatment centers, at Libreville, the capital city of Gabon and at Oyem and Koulamoutou, two rural cities between March 2015 and June 2016. P. falciparum-infected HIV adults were selected. Analysis of Pfdhfr and Pfdhps genes was performed using high resolution melting (HRM) technique. Results. Pfdhps A581G mutation was found in 23.5% (8/34) of the isolates. Triple Pfdhfr mutation (51I-59R-108N) was predominant (29.4%; n=10) while 17.6% (n=6) of the isolates carried a quadruple mutation (Pfdhfr 51I-59R-108N + Pfdhps 437G; Pfdhfr 51I-108N + Pfdhps 437G-Pfdhps581G; Pfdhfr 51I-59R-108N + Pfdhps 581G). Highly resistant genotype was detected in around 10% (n=3) of the isolates. The quintuple mutation (triple Pfdhfr 51I-59R-108N and double Pfdhps437-581) was only found in isolates from two patients who did not use CTX. The most frequent haplotypes were those with a single mutation (NCNIAKA) (36%) and a quadruple mutation (NCIIGKG, NRIIGKA, and NRIIAKG). Mixed unknown genotypes were found at codon 164 in three isolates. Mixed genotypes were more frequent at codons 51 (23.5%; n=8) and 59 (20.5%; n=7) (p<0.01). Conclusion. Pfdhps A581G mutation as well as new combination of quintuple mutations is found for the first time in isolates from HIV-infected patients in Gabon in comparison to a previous study. The detection of these genotypes at a nonnegligible frequency underlines the need of a regular surveillance of antifolates drug resistance.


2021 ◽  
Vol 10 (1) ◽  
Author(s):  
Tobias O. Apinjoh ◽  
Veronica N. Ntasin ◽  
Phil Collins C. Tataw ◽  
Vincent N. Ntui ◽  
Dieudonne L. Njimoh ◽  
...  

Abstract Background Malaria remains a significant health challenge in sub-Saharan Africa, with early diagnosis critical to reducing its morbidity and mortality. Despite the increasing Plasmodium spp. diagnostic capabilities, access to testing is limited in some cases by the almost absolute requirement for blood from potentially infected subjects as the only sample source for all conventional methods. A rapid test on non-invasive specimen with comparable performance to microscopy for the screening or diagnosis of all participants is invaluable. This study sought to compare conventional and non-invasive diagnostic tools for detecting Plasmodium falciparum. Methods This was a cross-sectional study, carried out between March and August 2019 to evaluate and compare the diagnostic performance of a PfHRP2/pLDH-based malaria rapid diagnostic test (mRDT) on patients’ blood, saliva and urine relative to conventional light microscopy and nested PCR at outpatient clinics in the Buea and Tiko health districts of Southwestern Cameroon. The significance of differences in proportions was explored using the Pearson’s χ2 test whereas differences in group means were assessed using analyses of variance. Results A total of 359 individuals of both sexes, aged 1–92 years, were enrolled into the study. Of the 301 individuals tested by light microscopy and mRDTs on blood, saliva and urine, 84 (27.9%), 81 (26.9%), 87 (28.9%) and 107 (35.5%) respectively were positive. However, only 34.3%, 90.5%, 91.4%, 83.9% and 65.4% febrile, light microscopy and mRDT positives on blood, saliva and urine respectively had P. falciparum infection as confirmed by PCR. The sensitivity and specificity of presumptive diagnosis, light microscopy and mRDT on blood, saliva and urine were 86.9% and 19.7%, 77.8% and 96.1%, 75.8% and 96.6%, 74.5% and 93.1%, and 70.7% and 81.8%, respectively. The agreement between mRDT on saliva (k = 0.696) and microscopy (k = 0.766) compared to PCR was good. Conclusion The study highlighted the low performance of presumptive diagnosis, reinforcing the need for parasitological tests prior to antimalarial therapy. The higher PfHRP2/pLDH mRDT parasite detection rates and sensitivity in saliva compared to urine suggests that the former is a practical adjunct to or alternative worth optimising for the routine diagnosis of malaria. Graphic Abstract Flow chart for diagnosis of P. falciparum infection by light microscopy, rapid diagnostic tests and nested PCR.


Author(s):  
Sangita Devubhai Vasava ◽  
Sucheta Jitendra Lakhani ◽  
Jitendra Devjibhai Lakhani

Introduction: There are several tropical mosquito borne infections, such as Malaria and Dengue, these are the two major and common arthropod borne infections that cause high morbidity and mortality in many patients and are major public health concern worldwide. Concurrent malaria and dengue infection is an important condition that is infrequently reported. Aim: To find out the prevalence rate of co-infection for both dengue and malaria and also to associate the severity of such co-infections with symptoms and haematological parameters. Materials and Methods: In observational cross-sectional study, samples were collected from 604 febrile patients clinically suspected for malaria and dengue attending the Medicine Out Patient Department (OPD) of Dhiraj Hospital, Gujarat, India. The samples were diagnosed for malaria by using rapid malaria antigen test kit and peripheral blood smear microscopy for the identification of Plasmodium spp. The dengue NS-1 antigen, (Immunoglobulin) IgM and IgG antibodies rapid kit were use to rule out dengue infection. The statistical analysis done by software EPI info. Results: In the present study, out of 604, 58 patients (9.6%) were positive for malaria and 80 patients (13.24%) were positive for dengue, while 21 patients (3.47%) were having concomitant infection with both dengue and malaria. The most affected age group was 31-60 years, 45.53%. The study revealed male preponderance (61.59%), hepatomegaly and jaundice were seen in 52.38% patients, haemorrhagic manifestations in 23.80%, kidney failure in 4.76% (1), and thrombocytopenia (platelet count <150,000/cubic millimeter (cmm)) were noted in 95.23% patients. Conclusion: It is mandatory to test any febrile patient for both malaria and dengue so that the diagnosis of the patient is not held back by missing any one. The clinicians who treat febrile patients in or from endemic areas must thoroughly examine and diagnose for both malaria and dengue, still one or the other is positive. Vector control, health education and good hygiene are community based preventive measures that are needed to control both the diseases.


2019 ◽  
Vol 11 (1) ◽  
Author(s):  
Shalala Zeynalova

ObjectiveSchmallenberg virus (SBV) is an orthobunyavirus that primarily infects domestic and wild ruminants and causes symptoms such as transient fever, diarrhea, reduced milk production, congenital malformations and abortion. The first virus was identified in 2011 at the onset of a major outbreak in Europe (Germany, Hungary, and France).IntroductionIn 2012 - 2017 in Azerbaijan there was an unexpected increase of abortions in cattle and sheep that was unrelated to brucellosis or chlamydia infection. The first confirmed case of Schmallenberg disease was received from Beylagan district of Azerbaijan in October 2012. The import of cattle from Europe to Azerbaijan has commenced in 2012. Therefore, the surveillance study was launched to determine spread of infection among cattle and sheep and to monitor the situation in the country.MethodsState Veterinary Control Service notified 42 Regional Veterinary Offices of Azerbaijan to commence the monitoring of Schmallenberg disease. Blood samples were collected from sheep, and cattle and biopsies of heads or necks from aborted fetuses were sampled too.. The collected samples were tested in the Republican Veterinary Laboratory. ELISA was used to investigate the presence of specific antibodies against Schmallenberg virus in the blood samples using IDEXX Schmallenberg Ab Test Kit. The commercially available real-time PCR kits (VetMAX™ Schmallenberg Virus Kit) were applied to test the biopsy samples. Both tests were recommended by the World Organization for Animal Health.ResultsTotal, 40,257 blood samples were collected from suspicious cattle and sheep. 671 biopsies samples were taken from fetuses. 4,281 cattle and 999 sheep with antibodies against SBVwere detected. The PCR results showed that the 77 biopsies samples were positive for SBV. The highest numbers of seropositive animals were found in Ganja, Aghdash, Barda, and Baku.ConclusionsThis biosurveillance study determined SBV in the samples of cattle and sheep in Azerbaijan, therefore, it is important to carry out annual seromonitoring and start the vaccination program. It is essential to check the passport of imported cattle, which has the disease history and seroprevalence of SBV.ReferencesLaloy, E., Breard, E., Sailleau, C., Viarouge, C., Desprat, A., Zientara, S., Klein, F., Hars, J., Rossi, S., 2014. Schmallenberg virus infection among red deer, France, 2010-2012. Emerg. Infect. Dis. 20, 131–134. https://doi.org/10.3201/eid2001.130411Larska, M., Krzysiak, M.K., Kesik-Maliszewska, J., Rola, J., 2014. Cross-sectional study of Schmallenberg virus seroprevalence in wild ruminants in Poland at the end of the vector season of 2013. BMC Vet. Res. 10, 967. https://doi.org/10.1186/s12917-014-0307-3 


PLoS ONE ◽  
2021 ◽  
Vol 16 (7) ◽  
pp. e0253953
Author(s):  
Getahun Asebe ◽  
Daniela Michlmayr ◽  
Gezahegne Mamo ◽  
Woldaregay Erku Abegaz ◽  
Adugna Endale ◽  
...  

Yellow fever (YF), Chikungunya (CHIK), and Zika(ZIK) are among re-emerging arboviral diseases of major public health concern. Despite the proximity of the Gambella Region to South Sudan where arboviral cases have been recorded repeatedly the current epidemiological situation is unclear in this part of southwest Ethiopia. Therefore, we conducted a community-based seroprevalence survey of YF virus (YFV), CHIK virus (CHIKV), and ZIK virus (ZIKV) infections in two selected districts. A cross-sectional study was conducted in two locations of the Gambella region (Lare and Itang) to investigate the seroprevalence of these viruses’ infections. Blood samples were collected from the study participants and screened for IgG antibodies specific to YFV and CHIKV infections using enzyme-linked immunosorbent assays (ELISA). For the detection of ZIKV specific IgG antibodies, Blockade-of-binding ELISA was used. Data were analyzed using the STATA version 13.1 Softwares. A total of 150 individuals (96 males and 54 females, age ranging from 18 to 65 years, mean age ± SD = 35.92 ± 10.99) participated and provided blood samples. Among the 150 samples 135, 90, and 150 were screened for YFV, CHIKV, and ZIKV, respectively. Hence, 2.9% (95% CI: 1.1–7.7%), 15.6% (95% CI: 9.3–24.8%), and 27.3% (95% CI: 20.7–35.3%) of samples tested positive for IgG antibodies to YFV, CHIKV, and ZIKV infections, respectively. Among the individual seropositive for ZIKV, YFV and CHIKV, only six, one and three had a history of residence outside the Gambella region respectively. Agro-pastoral occupation was significantly associated with a higher prevalence of IgG against CHIKV (AOR = 14.17; 95%CI: 2.30, 87.30) and residency in the Lare district (AOR = 11; 95%CI: 3.31, 39.81) was found to be significantly associated with a higher prevalence of IgG against ZIKV. Our findings revealed the occurrence of YFV, CHIKV and ZIKV infections in the study locations.


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