Genetyping Of Plasmodium Falciparum In Yunnan Province, China And Study Of Drug-resistant Gene Pfcrt~*

【Objective】Genetyping of Plasmodium falciparum in Yunnan Province,China-Myanmar border district,using molecular biology technology.Understanding the distribution of the alleic genotye.And studing of drug-resistant gene pfcrt.【Methods】Parasite genome DNA was extracted from 100 samples of blood the filter papers or Plasmodium falciparum in vitro culture.Type the Merozotite Surface Protein 1 (MSP1),Merozotite Surface Protein 2(MSP2) and glutamate rich protein(GLURP) by nested PCR.And genotype the Plasmodium falciparum isolated from blood these samples. Selected the strains of a single infection for drug-resistant Plasmodium falciparum in vitro studies,and gene polymorphism analysis.At the same time carried out nested polymerase chain reaction(nested-PCR),examined the mutation at 76 codon of the P.falciparum chloroquine resistance transporter(pfcrt) gene by using a similar PCR-restriction fragment length polymorphism(PCR-RFLP) method.Observation of the existence of mutations. Finally the results were statistically analyzed.【Results】(1)MSP 1 genotypes:99 blood samples,MAD20-type,Kl-type and RO33-type alleles were obtained in 53(53.50%),47(47.50%) and 20(20.20%) samples respectively. Both MAD20-type and KI-type alleles in 20(20.20%)samples.All of the three type alleles in 4(4.04%) samples.(2) MSP2 genotypes:99 blood samples,3D7-type and FC27-type alleles were obtained in 45(45.50%),48(48.50%) samples respectively and both alleles in 13(13.10 %) samples.(3) GLURP genotypes:99 blood samples single infections in 56(56.60%) samples.Sizes of the PCR products approximately at 500-1000 bp.While only the size of 750 bp was raised frequently.For the pfcrt code 76 mutation was 59/60(98.33%) in Yunnan Procince.【Conclusions】Plasmodium falciparum in Yunan Province of MSP 1 and MSP2 genes, such as living with MAD20-type alleles,respectively,and FC27-type was the dominant genotype;The mixed infection MAD20 and K1 allelic types of MSP1 were obtained in 20, K1-type,MAD20 and RO33 type is a mixed infection of 4.The mixed genotypes of MSP2 were 13 infections.And two genes MSP1 and MSP2 mixed infection at the same time,only four of the samples.MSP1,MSP2 and GLURP genes of mixed infection was 0.Different alleles of the mixed-type infection is very low.GLURP gene size of 750 bp was raised frequently.