Obeserving On Genetic Variability And Isoenzyme Patterns Of Ten Strains Of Blastocystis Hominis From Guangxi

Objective:To analyze the genotypes of ten strains of Blastocystis hominis(Bh) from Guangxi. And to observe malate dehydrogenase (MDH, EC 1.1.1.37),alkaline phophatase(ALP, EC.3.1.3.1,Lactate dehydrogenase (LDH, EC.1.1.1.27) and Esterase(EST, EC.3.1.1.1) patterns of the ten strains for the classification of Blastocystis hominis.Methods:Blastocystis hominis agents were obtained from patients's fecal specimen and cultivated in vitro. Genomic DNA of Blastocystis hominis was extracted, and to identify genotypes by PCR using seven pairs of known sequenced-tagged site (STS) primers. All Blastocystis hominis strains were made electrophoretic samples that were separated by means of polyaerylamide gel slab electronphoresis(PAGE). Then stained 4 enzymes specific stainirespectively and isoenzyme bands were record with relative mobility (Rm).Results:Eight of ten strains were identified as genotype 1 by PCR amplification with the primes SB83. Two strains unamplified with any STS primes were unknown genotypes.There were 7 enzyme bands in MDH patterns from ten Bh. Rm34, Rm47, Rm51,Rm55 and Rm59 were hackneyed.Rm34 and Rm51 enzyme bands were entirely identical in ten Bh; 5 enzyme bands existed in ALP patterns, Rm22,Rm25,Rm28,Rm35 and Rm38; 10 enzyme bands in LDH patterns from ten Bh. Rm37and Rm92 were hackneyed; 12 enzyme bands existed in EST patterns.Conclusions:By studying the genotypes of ten strains of Blastocystis hominis from Guangxi, we found there exist genotype 1 and unknown genotypes in Guangxi.Isoenzyme patterns may indicate genetic difference among Bh. And the results of genotype analysis and isoenzyme patterns are uniform. So isoenzyme patterns can offer potent evidence for genotyping of Blastocystis.hominis.Isoenzyme patterns can disclose the genetic difference in the same genotype more sensitively. This study indicates there may exist sub-genotypes in genotype 1. So isoenzyme patterns play an important function on the study of Bh's taxology.