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PCR-RFLP Based genetic diversity of Plasmodium vivax genotypes in district Mardan, Pakistan / Diversidade genética baseada em PCR-RFLP de genótipos de Plasmodium vivax no distrito de Mardan, Paquistão
Ullah, I; Afridi, S G; Khan, A U; Israr, M; Ali, A; Shams, S; Jabeen, H; Rasool, A; Akbar, F; Rahat, M A; Haris, M; Khan, A; Siraj, M; Shah, M.
  • Ullah, I; Abdul Wali Khan University Mardan. Department of Biochemistry. Mardan. PK
  • Afridi, S G; Abdul Wali Khan University Mardan. Department of Biochemistry. Mardan. PK
  • Khan, A U; Abdul Wali Khan University Mardan. Department of Biochemistry. Mardan. PK
  • Israr, M; University of Swat. Department of Forensic Sciences. Swat. PK
  • Ali, A; Saidu Medical College. Department of Biochemistry. Saidu Sharif. PK
  • Shams, S; Abdul Wali Khan University Mardan. Department of Biochemistry. Mardan. PK
  • Jabeen, H; Women University Mardan. Department of Microbiology. Mardan. PK
  • Rasool, A; University of Swat. Centre for Biotechnology & Microbiology. Swat. PK
  • Akbar, F; University of Swat. Centre for Biotechnology & Microbiology. Swat. PK
  • Rahat, M A; Hazara University Mansehra. Department of Genetics. PK
  • Haris, M; Hazara University Mansehra. Department of Genetics. PK
  • Khan, A; Hazara University Mansehra. Department of Genetics. PK
  • Siraj, M; Abbottabad University of Science & Technology. Department of Zoology. Abbottabad. PK
  • Shah, M; University of Swat. Centre for Animal Sciences & Fisheries. Swat. PK
Braz. j. biol ; 82: e241110, 2022. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1278500
ABSTRACT
Plasmodium vivax is the most common human malaria parasite in Asian countries including Pakistan. Present study was designed to explore the genetic diversity of plasmodium vivax genotypes based on Pvmsp-3α and Pvmsp-3ßgenes using allelic specific nested PCR and RFLP assays markers from field isolates in district Mardan, Pakistan. Blood samples of 200 P. vivax malarial patients were collected after taking their written informed consent. Genetic diversity in nested PCR products was determined by Restriction Fragment Length Polymorphism (RFLP) utilizing Alu1 and PstI restriction enzymes for alpha and beta gene products digestion, respectively. For analysis the genetic diversity of the sub allelic variants of Pvmsp3α and Pvmsp3ß genes, Chi-Square test was performed by utilizing Minitab programming software 18. The P value 0.05 was considered as statistically significant. For Pvmsp3α genes after gel electrophoresis of digested products, four distinct genotypes were obtained from total of 50 samples; type A 35 (70%) (1.5-2.0 kb), 12 of type B (24%) (1.5-1.7 kb), 2 of type C (4%) (0.5-1.5) and one for type D (2%) (0.5-0.65 kb) which could be characterized into 9 allelic pattern (A1-A4, B1-B3, C1, D), in which A3 remained the most predominant. For Pvmsp-3ßgenes, three distinct genotypes were obtained from 50 samples; 40(80%) of type A (1.5-2.5 kb), 9 (18%) of type B (1.0-1.5kb) and 1(2%) of type C (0.65 kb) which could be characterized into 6 allelic patterns (A1-A3, B1-B2, and C1). Most dominant one in Type A was A1 alleles which were noted (46%), while in Type B, the most dominant were B1 (10%).This study is the first ever report of molecular epidemiology and genetic variation in Pvmsp-3α and Pvmsp-3ß genes of P. vivax isolates by using PCR/RFLP from District Mardan and showed a remarkable level of genetic diversity in the studied genes of circulating parasites in the study area. The results of this study will contribute in future studies about the genetic structure of parasite and vaccine development against the malaria.
RESUMO
O Plasmodium vivax é o parasita da malária humana mais comum nos países asiáticos, incluindo o Paquistão. O presente estudo foi desenhado para explorar a diversidade genética de genótipos de Plasmodium vivax baseados nos genes Pvmsp-3α e Pvmsp-3ß, usando marcadores de ensaios alélicos nested PCR e RFLP de isolados de campo no distrito de Mardan, Paquistão. Amostras de sangue de 200 pacientes com malária por P. vivax foram coletadas após assinatura do termo de consentimento livre e esclarecido. A diversidade genética em produtos de PCR nested foi determinada por polimorfismo de fragmento de restrição (RFLP) utilizando as enzimas de restrição Alu1 e PstI para a digestão dos produtos dos genes alfa e beta, respectivamente. Para análise da diversidade genética das variantes subalélicas dos genes Pvmsp3α e Pvmsp3ß, o teste Qui-quadrado foi realizado utilizando o software de programação Minitab 18. O valor P = 0,05 foi considerado estatisticamente significativo. Para os genes Pvmsp3α, após eletroforese em gel de produtos digeridos, quatro genótipos distintos foram obtidos de um total de 50 amostras; tipo A 35 (70%) (1,5-2,0 kb), 12 do tipo B (24%) (1,5-1,7 kb), 2 do tipo C (4%) (0,5-1,5) e um para o tipo D (2%) (0,5-0,65 kb), que podem ser caracterizados em nove padrões alélicos (A1-A4, B1-B3, C1, D), em que A3 permaneceu como o mais predominante. Para Pvmsp-3ßgenes, três genótipos distintos foram obtidos a partir de 50 amostras; 40 (80%) do tipo A (1,5-2,5 kb), 9 (18%) do tipo B (1,0-1,5 kb) e 1 (2%) do tipo C (0,65 kb), que podem ser caracterizados em seis padrões alélicos (A1-A3, B1-B2 e C1). Os mais dominantes no tipo A foram o alelo A1, observados em 46%, enquanto, no tipo B, os mais dominantes foram B1 (10%). Este estudo é o primeiro relato de epidemiologia molecular e variação genética em Pvmsp-3α. Os genes Pvmsp-3ß de isolados de P. vivax utilizando PCR/RFLP do Distrito Mardan mostraram um nível notável de diversidade genética nos genes estudados de parasitas circulantes na área de estudo. Os resultados desse estudo contribuirão em estudos futuros sobre a estrutura genética do parasita e o desenvolvimento de vacinas contra a malária.

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Full text: Available Index: LILACS (Americas) Main subject: Plasmodium vivax / Protozoan Proteins Limits: Humans Country/Region as subject: Asia Language: English Journal: Braz. j. biol Year: 2022 Type: Article Institution/Affiliation country: Abbottabad University of Science & Technology/PK / Abdul Wali Khan University Mardan/PK / Hazara University Mansehra/PK / Saidu Medical College/PK / University of Swat/PK / Women University Mardan/PK

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Full text: Available Index: LILACS (Americas) Main subject: Plasmodium vivax / Protozoan Proteins Limits: Humans Country/Region as subject: Asia Language: English Journal: Braz. j. biol Year: 2022 Type: Article Institution/Affiliation country: Abbottabad University of Science & Technology/PK / Abdul Wali Khan University Mardan/PK / Hazara University Mansehra/PK / Saidu Medical College/PK / University of Swat/PK / Women University Mardan/PK