Academic journal article Iranian Journal of Public Health

Optimization of Real Time Pcr for Detection of Brucell Abortus

Academic journal article Iranian Journal of Public Health

Optimization of Real Time Pcr for Detection of Brucell Abortus

Article excerpt

Background: Brucellosis is a zoonotic disease transferred from infected animal to human. Six classic species involved B. abortus, B. melitensis, B. suis, B.canis, B.neotoma and B. ovis. In recent years four new species consist of B.ceti, B. maris, B. pinipedialis and B.inopinata were reported. Standard and classic methods are time consuming and have low specificity and sensitivity.Several Molcular methods have been developed but many of them are not able to detect all Brucella intraspe- cies biovars, thus developing fast and specific techniques for identification of brucellosis is essential.

Methods: Bases on bioinformatic study, B. abortus specific positions were found then a Real time PCR assay was de- signed. Bacterial samples used in this study consist of Brucella standard, vaccinal and 15 filed strains isolated from cow milk. High Pure PCR Template preparation Kit was used for DNA extraction. Sensitivity and specificity assays were performed

Results: Real time PCR was 100% identical with conven- tional bacteriological method and phage typing assay. Moreo- ver,based on sensitivity testes this presented real time PCR method had detection limit 400 fg B. abortus strain 544 tem- plate DNA.

Conclusion: Brucellosis detection by classic methods con- sists of bacterial isolation and serological assays that have sensitivity between 15 to 70 percent. …

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