Genetic Diversity And Multiplicity Of Infections Of Plasmodium Falciparum Clinical Isolates From Adama And Its Surroundings, Oromia, Ethiopia

Abstract

Genetic Diversity Of P. Falciparum Can Be Determined By Genotyping Of The Polymorphic Regions Like Block 2 Region Of Merozoite Surface Protein (Msp-1) And Block-3 Region Of Merozoite Surface Protein (Msp-2). Understanding Of These Polymorphic Genes Of P. Falciparum Can Inform Mechanisms Underlining Disease Pathology, Identification Of The Circulating Parasite Clones, Transmission Intensity And Potential Deficiencies And/Or Strengths Of The Ongoing Malaria Control And Elimination Efforts In The Locality. Detailed Understanding Of Local Genetic Polymorphism Is An Input To Pave The Way For Better Management, Control And Elimination Of Malaria. The Aim Of This Study Was Therefore To Explore The Status Of These Polymorphic Genes And Their Implications In Adama And Its Surrounding. For This Purpose, A Total Of 171 Dbs Sample From Uncomplicated P. Falciparum Patients Was Collected From Adama, Modjo, Wonji, Awash Malkasa, And Olanciti, Through September 2019 To August 2020. From These, 139 And 148 Clinical Isolates Were Successfully Amplified For Msp-1 And Msp-2 Genes, Respectively, Using Specific Primers. Nested Pcr Amplification Was Conducted Targeting K1, Mad20, And Ro33 Alleles For Msp-1 And 3d Andfc27 Alleles For Msp-2 Gene Followed By Gel Electrophoresis For Fragment Analysis. Based On The Detection Of A Pcr Fragment, Infections Were Classified As Monoclonal Or Multiple Infections. Accordingly, 19 Different Size Polymorphism Of Msp-1 Gene Were Identified In This Study, With 67(48.2 %) Mad20, 18 (13 %) K-1 And 18 (13 %) Ro33 Allelic Family. Whereas, The Multiple Infections Were 21(15 %), 8 (5.8 %), 4(2.9 %), 3(2.2 %) For Mad20+K-1, Mad20+Ro33, K-1+ Ro33, And Mad20+K-1+ Ro33, Respectively. The Overall Multiplicity Of Infection (Moi) Was 1.42 And The Expected Heterozygosity (He) Was 0.65. Moreover, 17 Different Polymorphic Forms Of Msp-2 Allelic Fragments Were Detected In The Study Area. A Total Of 47 (31.8%) And 41(27.7%) Were Detected For 3d7 And Fc27 Allelic Family, Respectively. Furthermore, For Msp-2 Gene: The Multi Clonal Allele Type Accounted For 60(40.5%). The Overall Moi And Expected Heterozygosity Were 1.46 And 0.68, Respectively. The Study Also Revealed Presence Of ??? 2 Clones Of Infection In Patient Isolates With The Same Allele In Both Markers (Msp-1 And Msp-2) Genes. Therefore, The Study Sheds Light On The Presence Of Moderate Level Of Genetic Diversity Of P. Falciparum In The Study Area,Compared With Similar Study In Different Parts Of Ethiopia, Demanding Further Scale Up Of The Ongoing Control And Elimination Efforts.