Diagnose of malaria infection with light microscopy and Nested- PCR methods and phylogenetic Analysis in peripheral blood expansions obtained from suspected patients in Hormozgan and Kerman provinces

Introduction and Objective: Malaria remains an important public health problem. This infection is a life-threatening disease caused by with Plasmodium protozoa. Rapid diagnostic tests (RDTs), for accurate and reliable diagnosing malaria infection is necessary. The purpose of this study was to diagnose of malaria infection with light microscopy (LM) and Nested- PCR methods and phylogenetic Analysis in peripheral blood expansions obtained from suspected patients in Hormozgan and Kerman provinces.Materials and Methods: 38 and 4 blood smears collected from Hormozgan and Kerman provinces, respectively. These blood smears were evaluated by light microscopy and Nested- PCR methods. Then, nine Plasmodium-positive samples were used to genetic diversity determination. Results: light microscopy results of Hormozgan province show that 4 P. falciparum, 33 p. vivax and 1 mixed infection with P. falciparum and P. vivax. For Kerman province, only 4 P. vivax was detected by LM. Nested PCR showed there is 2 P. falciparum, 31 p. vivax and 5 mixed infection with P. falciparum and P. vivax in Hormozgan province. In addition, Nested PCR confirmed that 100% (n=4) in Kerman province is P. vivax. Phylogenic results showed that the isolates H19, H41, H42, H33, A plu 3 and B plu 3 belong to Hormozgan province and isolates f K2, K3 and K4 belong to Kerman province, respectively. Isolate H33 located in subclade I and isolates H19, H42, and H41 were in sub clade II. Isolates of K 2, K3 and K4 placed in sub clade III. All of subclade I to III were p. vivax. Subclade IV involved A plu 3 and B plu 3 isolates and were p. falciparum. Conclusion: Use of rapid diagnostic tests such as Nested- PCR for accurate and reliable diagnosing malaria infection especially in endemic areas are recommended.

کلیدواژه ها:

Malaria ، Light microscopy ، Nested PCR ، Plasmodium falciparum ، Plasmodium vivax.