Validation of real-time PCR technique for detection of Mycobacterium bovis and Brucella abortus in bovine raw milk

Mascarenhas, Débora Rocha; Schwarz, David Germano Gonçalves; Júnior, Antônio Augusto Fonseca; Oliveira, Tatiana Flávia Pinheiro de; Moreira, Maria Aparecida Scatamburlo

doi:10.1007/s42770-020-00319-9

Cited by 3 publications

(2 citation statements)

References 20 publications

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“…To date, no tests that can help differentiate between vaccinated and naturally infected individuals have been officially approved. At present, a series of molecular methods was employed for the early and rapid diagnosis of infectious diseases, such as quantitative polymerase chain reaction [ 38 ]. Moreover, next-generation sequencing holds the potential for improving clinical and public health microbiology [ 39 ], and laboratories may be able to replace many traditional microbiology processes with a single workflow that accommodates a wide array of pathogens [ 40 ], which will prevent occupational Brucella spp.…”

Section: Discussionmentioning

confidence: 99%

A systematic analysis of and recommendations for public health events involving brucellosis from 2006 to 2019 in China

Liu

Wang²,

Tian³

et al. 2022

Annals of Medicine

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Brucellosis is a severe public health problem in China. However, analysis on related infection events is lacking. We performed a systematic analysis of brucellosis laboratory infection and vaccine infection events from 2006 to 2019 in China based on the published literatures. Our analysis showed that most laboratory Brucella infections in hospitals were found in Southern China. The identification and handling of suspected samples of Brucella infection without following the recommended biosafety protection was the main risk factor. It is important to strengthen the preventive awareness of clinical laboratory staff and physicians, while highlighting the compulsory handling and identification of suspected Brucella -infected samples in biosafety facilities and following biosafety practices. However, a severe Brucella infection accident at the Northeast Agricultural University, with 28 positive cases, showed that strengthening the management in teaching experiments of students in the veterinary-related profession is essential. However, cluster S2 vaccine strain infection events caused by vaccination and production were mainly observed in Northern China. Strengthening vaccination skills, personal protection, and improving the biosafety management of vaccine production and implementing regular risk surveillance is mandatory. Our analysis provides helpful clues for control of public health events involving brucellosis, as well as implementing intervention strategies is urgent.

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Section: Discussionmentioning

confidence: 99%

A systematic analysis of and recommendations for public health events involving brucellosis from 2006 to 2019 in China

Liu

Wang²,

Tian³

et al. 2022

Annals of Medicine

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“…However, PCR requires adequately trained personnel along with costly reagents and equipment [ 11 ]. Furthermore, calcium may inhibit polymerase activity by competing with magnesium ions present in milk [ 12 ]. During an outbreak, if a cow is determined to be negative, there is no suggestion to perform further testing.…”

Section: Introductionmentioning

confidence: 99%

High-Performance Detection of Mycobacterium bovis in Milk Using Recombinase-Aided Amplification–Clustered Regularly Interspaced Short Palindromic Repeat–Cas13a–Lateral Flow Detection

Wang,

et al. 2024

Foods

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Mycobacterium bovis (M. bovis), the microorganism responsible for bovine tuberculosis (bTB), is transferred to people by the ingestion of unpasteurized milk and unprocessed fermented milk products obtained from animals with the infection. The identification of M. bovis in milk samples is of the utmost importance to successfully prevent zoonotic diseases and maintain food safety. This study presents a comprehensive description of a highly efficient molecular test utilizing recombinase-aided amplification (RPA)–clustered regularly interspaced short palindromic repeat (CRISPR)-associated protein (Cas) 13a–lateral flow detection (LFD) for M. bovis detection. In contrast to ELISA, RPA–CRISPR–Cas13a–LFD exhibited greater accuracy and sensitivity in the detection of M. bovis in milk, presenting a detection limit of 2 × 100 copies/μL within a 2 h time frame. The two tests exhibited a moderate level of agreement, as shown by a kappa value of 0.452 (95%CI: 0.287–0.617, p < 0.001). RPA–CRISPR–Cas13a–LFD holds significant potential as a robust platform for pathogen detection in complex samples, thereby enabling the more dependable regulation of food safety examination, epidemiology research, and medical diagnosis.

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Recovery of Brucella in raw milk Minas artisanal cheese approved for consumption by official inspection agency in Brazil: assessment of prevalence and risk factors through One Health integrated approaches

Silva

Duch

Lage

et al. 2022

Transactions of the Royal Society of Tropical Medicine and Hygiene

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Background Minas artisanal cheese (MAC) from the Serro region is a Brazilian intangible cultural heritage. Produced from raw milk, it may carry zoonotic pathogens such as Brucella. This study included a randomized survey for the prevalence of Brucella-positive MAC and its associated factors. Methods MAC samples (n=55), each one from a different rural family-based cheese-processing agroindustry, were analysed for Brucella by direct polymerase chain reaction (PCR) species-specific DNA detection and cultivation-based approaches. Results Among 55 MACs that were analysed, we found 17 Brucella DNA-positive samples (30.9% [95% confidence interval {CI} 18.7 to 43.1]) by PCR and, for the first time, from one MAC (1.8% [95% CI 0.5 to 9.7]), viable Brucella abortus was recovered by cultivation. Higher values for two variables, the number of lactating cows per herd (p=0.043) and daily milk production per herd (p=0.043), were each associated with Brucella-positive MAC, which concentrated in three high-risk and one low-risk spatial clusters. Conclusions MAC may be a source of Brucella for humans, since the positive samples were from batches that were sold by cheesemakers. This should be of concern and encourage cooperation between the health and agriculture sectors in order to mitigate this public health risk through One Health integrated approaches.

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Validation of real-time PCR technique for detection of Mycobacterium bovis and Brucella abortus in bovine raw milk

Cited by 3 publications

References 20 publications

A systematic analysis of and recommendations for public health events involving brucellosis from 2006 to 2019 in China

A systematic analysis of and recommendations for public health events involving brucellosis from 2006 to 2019 in China

High-Performance Detection of Mycobacterium bovis in Milk Using Recombinase-Aided Amplification–Clustered Regularly Interspaced Short Palindromic Repeat–Cas13a–Lateral Flow Detection

Recovery of Brucella in raw milk Minas artisanal cheese approved for consumption by official inspection agency in Brazil: assessment of prevalence and risk factors through One Health integrated approaches

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