Genetic markers of resistance to pyrimethamine and sulfonamides in Plasmodium falciparum parasites compared with the resistance patterns in isolates of Escherichia coli from the same children in Guinea-Bissau

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.

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Detection of virulence factors and antimicrobial resistance patterns in shiga toxin-producing Escherichia coli isolates from sheep

Pesquisa Veterinária Brasileira ◽

10.1590/s0100-736x2015000900002 ◽

2015 ◽

Vol 35 (9) ◽

pp. 775-780 ◽

Cited By ~ 5

Author(s):

Marcos R.A. Ferreira ◽

Talícia dos S. Silva ◽

Ariel E. Stella ◽

Fabricio R. Conceição ◽

Edésio F. dos Reis ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Virulence Factors ◽

Shiga Toxin ◽

Resistance Profile ◽

Antimicrobial Resistance Profile ◽

Healthy Sheep ◽

Resistance Patterns ◽

Group A ◽

Stx1 Gene

Abstract: In order to detect virulence factors in Shiga toxin-producing Escherichia coli (STEC) isolates and investigate the antimicrobial resistance profile, rectal swabs were collected from healthy sheep of the races Santa Inês and Dorper. Of the 115 E. coli isolates obtained, 78.3% (90/115) were characterized as STEC, of which 52.2% (47/90) carried stx1 gene, 33.3% (30/90) stx2 and 14.5% (13/90) both genes. In search of virulence factors, 47.7% and 32.2% of the isolates carried the genes saa and cnf1. According to the analysis of the antimicrobial resistance profile, 83.3% (75/90) were resistant to at least one of the antibiotics tested. In phylogenetic classification grouped 24.4% (22/90) in group D (pathogenic), 32.2% (29/90) in group B1 (commensal) and 43.3% (39/90) in group A (commensal). The presence of several virulence factors as well as the high number of multiresistant isolates found in this study support the statement that sheep are potential carriers of pathogens threatening public health.

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Production of recombinant proteins from Plasmodium falciparum in Escherichia coli

Biomédica ◽

10.7705/biomedica.v36i3.3011 ◽

2016 ◽

Vol 36 ◽

Cited By ~ 4

Author(s):

Ángela Patricia Guerra ◽

Eliana Patricia Calvo ◽

Moisés Wasserman ◽

Jacqueline Chaparro-Olaya

Keyword(s):

Escherichia Coli ◽

Plasmodium Falciparum ◽

Recombinant Proteins

Introducción. La producción de proteínas recombinantes es fundamental para el estudio funcional de proteínas de Plasmodium falciparum. Sin embargo, las proteínas recombinantes de P. falciparum están entre las más difíciles de expresar y cuando lo hacen usualmente se agregan dentro de cuerpos de inclusión insolubles.Objetivo. Evaluar la producción de cuatro proteínas de P. falciparum, usando como sistema de expresión dos cepas de Escherichia coli genéticamente modificadas para favorecer la producción de proteínas heterólogas y establecer una reserva de proteínas recombinantes puras y solubles y producir anticuerpos policlonales a partir de ellas.Materiales y métodos. Las proteínas recombinantes, las cuales correspondían a secuencias parciales de PfMyoA (Miosina-A) y PfGAP50 (proteína-asociada a glideosoma-50 kDa) y a las secuencias completas de PfMTIP (proteína de interacción con Miosina-A) y PfGAP45 (proteína asociada a glideosoma-45 kDa), fueron expresadas como proteínas de fusión con GST y luego purificadas y usadas para producir anticuerpos policlonales en ratón.Resultados. La expresión de las proteínas recombinantes fue mucho más eficiente en la cepa BL21-CodonPlus (la cual expresa tRNAs escasos en las bacterias silvestres), que en la cepa BL21-pG-KJE8. En contraste, aunque la cepa BL21-pG-KJE sobreexpresa chaperonas, no redujo la formación de cuerpos de inclusión. Conclusión. El uso de cepas de E. coli genéticamente modificadas fue fundamental para alcanzar altos niveles de expresión de las cuatro proteínas recombinantes evaluadas y permitió obtener dos de ellas en forma soluble. La estrategia utilizada permitió expresar cuatro proteínas recombinantes de P. falciparum en cantidad suficiente para inmunizar ratones y producir anticuerpos policlonales, y además conservar proteína pura y soluble de dos de ellas, para ensayos futuros.

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