Evaluation of a new set of Real-Time PCR for Brucella detection within human and animal samples

Document Type : Original Article

Abstract

A quantitative TaqMn Real-Time PCR assay was developed and its diagnostic value on human serum and livestock samples were evaluated. Brucella species could be distributed through communities as a biological agent. Rapid detection of biological threat agents is critical for timely therapeutic administration. Quantitative real-time PCR provides a rapid, sensitive and specific tool for molecular identification of this agent. We evaluated a new real-time PCR set by numerous human and livestock samples. For primers and probe designation BCSP31 of B. melitensis 16 M was used. Different sero-positive human samples and tissue samples livestock with suspected brucellosis were then assayed after primary adaptation of Q-PCR in the laboratory. The detection limit of the assay was 10 fg (equivalent to 2 genome). Totally 50 samples (25 sero-positive, 25 sero-negative) of patients and 75 samples from slaughtered livestock (due to brucellosis) were collected and assayed by Q-PCR, along with control samples. Brucella genome were detected by this real-time set from nearly 42% (5 out of 12) of negative samples by conventional PCR. This system also detected brucella contamination in 46 out of 75 animal samples.

Keywords

• Real-time PCR
• TaqMn
• brucella
• BCSP31

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