MLVA and MLST typing of Brucella from Qinghai,China

Background:The Qinghai-Tibet Plateau(QTP)of China is an extensive pastoral and semi-pastoral area,and because of poverty and bad hygiene conditions,Brucella is highly prevalent in this region.In order to adequately prevent this disease in the QTP region it is important to determine the identity of Brucella species that caused the infection.Methods:A total of 65 Brucella isolates were obtained from human,livestock and wild animals in Qinghai,a Chinese province in east of the QTP.Two molecular typing methods,MLVA(multi-locus variable-number tandem-repeat analysis)and MLST(multi locus sequence typing)were used to identify the species and genotypes of these isolates.Findings:Both MLVA and MLST typing methods classified the 65 isolates into three species,B.melitensis,B.abortus and B.suis,which included 60,4 and 1 isolates respectively.The MLVA method uniquely detected 34(Bm01~Bm34),3(Ba01~Ba03),and 1(Bs01)MLVA-16 genotypes for B.melitensis,B.abortus and B.suis,respectively.However,none of these genotypes exactly matched any of the genotypes in the Brucella2012 MLVA database.The MLST method identified five known ST types:ST7 and ST8(B.melitensis),ST2 and ST5(B.abortus),and ST14(B.suis).We also detected a strain with a mutant type(3-2-3-2-?-5-3-8-2)of ST8(3-2-3-2-1-5-3-8-2).Extensive genotype-sharing events could be observed among isolates from different host species.Conclusions:There were at least three Brucella(B.melitensis,B.abortus and B.suis)species in Qinghai,of which B.melitensis was the predominant species in the area examined.The Brucella population in Qinghai was very different from other regions of the world,possibly owing to the unique geographical characteristics such as extremely high altitude in QTP.There were extensive genotype-sharing events between isolates obtained from humans and other animals.Yaks,sheep and blue sheep were important zoonotic reservoirs of brucellosis causing species found in humans.