Abstract
BackgroundThe present study aimed to direct detect Mycobacterium bovis in milk (n = 401) and blood (n = 401) samples collected from 401 dairy cows of 20 properties located in the state of Pernambuco, Brazil, by real-time quantitative PCR (qPCR) targeting the region of difference 4 (RD4). Risk factors possibly associated with bovine tuberculosis (BTB) were also evaluated.ResultsOf the 802 samples analyzed, one milk (0.25 %) and eight blood (2 %) samples were positive for M. bovis in the qPCR and their identities were confirmed by sequencing. Animals positive for M. bovis were found in six (30 %) of the 20 properties visited. None of the risk factors evaluated were statistically associated with BTB.ConclusionsM. bovis DNA was detected in one milk sample what may pose a risk to public health because raw milk is commonly consumed in Brazil.
Highlights
The present study aimed to direct detect Mycobacterium bovis in milk (n = 401) and blood (n = 401) samples collected from 401 dairy cows of 20 properties located in the state of Pernambuco, Brazil, by real-time quantitative Polymerase Chain Reaction (PCR) targeting the region of difference 4 (RD4)
Quantitative real time PCR was performed using the same primer set used for amplification of the RD4 fragment of the positive control and a fluorescent probe that discriminates M. bovis from other M. tuberculosis complex members since it hybridizes with both the 5′ and 3′ RD4 deletion flanking sequences, which only occur directly adjacent to each other in M. bovis
Of the 802 samples analyzed, one milk (0.25 %) and eight blood (2 %) samples were positive for M. bovis in the quantitative PCR (qPCR) and their identities were confirmed by sequencing
Summary
Bovine tuberculosis (BTB) is caused by Mycobacterium bovis, a member of the Mycobacterium tuberculosis complex that affects mammals, including humans [1]. The tuberculin skin test is the diagnostic method recommended by the PNCETB and it must be followed by. Molecular techniques such as Polymerase Chain Reaction (PCR) have been used for BTB diagnosis in several clinical samples such as blood, milk and nasal exudates [2]. Standardization of direct methods for detection of M. bovis in clinical samples will enable a more accurate BTB diagnosis and facilitate epidemiological studies on M. bovis prevalence [1, 8]. We used qPCR for direct detection of Mycobacterium bovis in milk and blood samples of cattle from the state of Pernambuco, Brazil
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