Haplotype and mutation analysis of drug resistance Pfcrt and Pfmdr1 gene of imported Plasmodium falciparum
XU Chao, WEI Qing-kuan, LI Jin, XIAO Ting, YIN Kun, KONG Xiang-li, WANG Yong-bin, CUI Yong, SUN Hui, ZHAO Gui-hua, ZHU Song, YAN Ge, HUANG Bing-cheng
Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Diseases,Shandong Provincial Reference Laboratory for Malaria Diagnosis, Jining 272033, China
In order to understand the drug resistance Pfcrt and Pfmdr1 gene haplotypes of Plasmodium falciparum imported currently in Shandong Province, we analyzed the mutant genotypes and their distribution situation. The blood samples were collected from malaria cases infected with P. falciparum deriving from Africa during 2011 to 2013 in Shandong Province. P. falciparum genomic DNA was extracted and primers were designed according to the Pfcrt and Pfmdr1 gene sequences. Nested PCR amplification and sequencing were performed, and sequence alignment was carried out by using bioinformatics software. The results showed that all of 68 P. falciparum samples imported were amplified and sequencing successfully at codon 72-76 in Pfcrt gene and codon 86, 1 042 and 1 246 in Pfmdr1 gene. And 69.12% of the samples were wild type CVMNK in Pfcrt gene and 30.88% were mutation types, including CVIET, CVIDT and mixed types, of which the CVIET haplotype was the most. The 69.12% of the samples were wild type NND in Pfmdr1 gene and 30.88% were mutation types including YND and mixed types. Among the six African source countries? samples, mutant gene types existed in all samples in both Pfcrt and Pfmdr1 gene except Pfcrt genotype from Guinea were all wild type. Five samples had mutation types in both pfcrt and pfmdr1 gene together. What could be drawn conclusion from the results was that mutant Pfcrt and Pfmdr1 haplotypes expressed diversification in the imported P. falciparum cases in Shandong Province, and the proportion of mutant genotypes were less than wild genotypes, which suggested that it was not appeared serious chloroquine-resistant of the imported P. falciparum prevalence in Shandong Province currently.
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2016, Vol. 32 
