Diagnosis of human brucellosis using AMOS PCR
Abstract number: p947
Yu W., Nielsen K., Grushina T., Meka-Mechenko T.
Numerous PCR-based assays have been developed for the identification of Brucella to improve diagnostic capabilities. For some purposes, the simple identification of Brucella is adequate. However, PCR with IPS1 and IPS2 primers is a reliable tool for rapid identification of Brucella species and its differentiation from closely related organisms. A fundamental question that arises during epidemiological investigations of outbreaks is whether the outbreak strain is genetically related to a proposed strain. Highly discriminating genetic markers for characterizing bacterial strains can help in clarifying the genetic relationships among strains. The Brucella AMOS (AbortusMelitensisOvisSuis)-PCR assay was previously developed to identify and differentiate specific Brucella species.
Material and methods:
Twenty strains of B. melitensis isolated from humans and animals using traditional techniques during four field trips to rural areas in two regions of Kazakhstan. Total 360 people were examined. B. melitensis biovars1, 2 and 3 were isolated in 18 cases out 130 individuals presumed to be infected with Brucella sp. Two B. melitensis biovar 2 were isolated from sheep milk. Twenty isolates were identified and tested by the conventional biochemical tests, PCR-based and Brucella AMOS PCR
Twenty isolates were identified and tested by the conventional biochemical tests, PCR-based and Brucella AMOS PCR. This included 6 isolates identified as B. melitensis biovar 1; 10-identified as B. melitensis biovar 2; 4-identified as B. melitensis biovar 3. The results of classical typing for 20 B. melitensis cultures isolated from humans and animals were confirmed at genetic level by PCR. The high specificity of this PCR-based assay was demonstrated by testing all of the biovars of twenty strains of B. melitensis. The Brucella AMOS PCR correctly identified each isolate as a B. melitensis. However due to AMOS, but not biological typing, genetic features of 2 strains of B. melitensis biovar1 and 2 have been found. Two B. melitensis biovar 1 and 2 have been isolated from the people with serological positive acute brucellosis (SAT: 1: 1600).
The PCR-based test and AMOS-PCR could be used as a complementary test to identify genus Brucella and differentiate specific Brucella species. However, to explain further genetic distinctions, fingerprinting is required.
|Session name:||XXIst ISTH Congress|
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