Genome-wide unique insertion sequences among five Brucella species and demonstration of differential identification of Brucella by multiplex PCR assay

Paul, Souvik; Peddayelachagiri, Bhavani V.; Gogoi, Madhurjya; Nagaraj, Sowmya; Ramlal, Shylaja; Balakrishna, K.; Batra, Harsh Vardhan

doi:10.1038/s41598-020-62472-3

Cited by 11 publications

(8 citation statements)

References 27 publications

(13 reference statements)

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“…Brucella abortus pathogen as an important infectious agent of bovine brucellosis cannot be ignored, especially in countries/regions dominated by animal husbandry ( Karthik et al, 2014 ; Rahdar et al, 2018 ; Paul et al, 2020 ). Although traditional culture, serology, and biochemical tests are commonly used to diagnose bovine brucellosis, these methods are time-consuming, low-sensitivity, or prone to cross-reaction with other organisms ( Sergueev et al, 2017 ).…”

Section: Discussionmentioning

confidence: 99%

Rapid, ultrasensitive, and highly specific identification of Brucella abortus utilizing multiple cross displacement amplification combined with a gold nanoparticles-based lateral flow biosensor

et al. 2022

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Brucella abortus (B. abortus) as an important infectious agent of bovine brucellosis cannot be ignored, especially in countries/regions dominated by animal husbandry. Thus, the development of an ultrasensitive and highly specific identification technique is an ideal strategy to control the transmission of bovine brucellosis. In this report, a novel detection protocol, which utilizes multiple cross displacement amplification (MCDA) combined with a gold nanoparticles-based lateral flow biosensor (AuNPs-LFB) targeting the BruAb2_0168 gene was successfully devised and established for the identification of B. abortus (termed B. abortus-MCDA-LFB). Ten specific primers containing engineered C1-FAM (carboxyfluorescein) and D1-biotin primers were designed according to the MCDA reaction mechanism. These genomic DNA extracted from various bacterial strains and whole blood samples were used to optimize and evaluate the B. abortus-MCDA-LFB assay. As a result, the optimal reaction conditions for the B. abortus-MCDA-LFB assay were 66°C for 40 min. The limit of detection of the B. abortus-MCDA-LFB was 10 fg/μl (~3 copies/μl) for genomic DNA extracted from pure cultures of B. abortus isolate. Meanwhile, the B. abortus-MCDA-LFB assay accurately identified all tested B. abortus strains, and there was no cross-reaction with non-B. abortus pathogens. Moreover, the detection workflow of the B. abortus-MCDA-LFB assay for whole blood samples can be completed within 70 min, and the cost of a single test is approximately 5.0 USD. Taken together, the B. abortus-MCDA-LFB assay is a visual, fast, ultrasensitive, low-cost, easy-to-operate, and highly specific detection method, which can be used as a rapid identification tool for B. abortus infections.

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Section: Discussionmentioning

confidence: 99%

Rapid, ultrasensitive, and highly specific identification of Brucella abortus utilizing multiple cross displacement amplification combined with a gold nanoparticles-based lateral flow biosensor

et al. 2022

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“…The genus Brucella includes twelve species: Brucella abortus, B. melitensis, B. suis, B. canis, B. ovis, B. neotomae, B. microti, B. pinnipedialis, B. ceti, B. inopinata, B. papionis, and B. vulpis [26]. Three of these species have been further classified into biovars, B. abortus with eight, three for B. melitensis and five for B. suis [8] [26]. A summary of the Brucella species, their biovars and their primary host are presented in Table 1.…”

Section: Etiologic Agents Of Brucellosismentioning

confidence: 99%

“…Brucellosis has been reported in 86 different countries worldwide with more Table 1. Brucella spp and their hosts [8] [26] [27] [30].…”

Section: Epidemiologymentioning

confidence: 99%

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A Review on Brucellosis in Cameroon: Diagnostic Approaches, Epidemiology and Risk Factors for Infection

Eko¹,

Esemu²,

Nota³

et al. 2022

AiM

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Brucellosis is a neglected tropical zoonotic disease that threatens the food production and public health sectors. It is of considerable animal welfare and economic importance and is underreported in most parts of the world, especially in developing countries like Cameroon. Brucellosis has been reported in cattle, other domestic animals and humans in Cameroon. The burden of the disease is unclear, and the awareness remains questionable. It became necessary for this review to be carried out to highlight the diagnostic approaches used to confirm brucellosis in animals and humans, disease epidemiology and risk factors for infection. So far, reports of brucellosis in previous studies have been based on serology only. Seroprevalence data of Brucella antibodies in animals indicate the risk of human brucellosis in Cameroon. However, few investigations have been undertaken on human brucellosis, considering the different epidemiological settings. There is no report or unsuccessful attempts to identify Brucella species circulating in Cameroon. It could largely be attributed to a lack of standard laboratories for testing and the lack of consumables. The way forward will require a surveillance system for brucellosis in the country, educating all sectors affected and drafting a diagnostic protocol for high-risk individuals.

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“…Brucellosis, caused by a group of bacteria of the genus Brucella, is a widespread, endemic, and neglected zoonotic disease in most low-income countries [1,2]. Te genus Brucella includes twelve species, some of which are hostspecifc [3,4]. Te disease afects both humans and domestic animals, particularly cattle [2,5].…”

Section: Introductionmentioning

confidence: 99%

Identification and Prevalence of Brucella Species Circulating among Cattle Slaughtered in the Douala and Buea Municipalities of Cameroon

Eko

Esemu

Njouendou

et al. 2023

Veterinary Medicine International

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Brucellosis is a neglected zoonotic disease affecting the livestock sector in low-income countries. Cameroon, a lower-middle-income country in sub-Saharan Africa, has reported the prevalence of brucellosis in regions where livestock rearing is the principal economic activity. However, the presence of the disease has not been reported in southern regions receiving cattle for consumption by their population. In addition, there is no report on the Brucella species circulating in Cameroon. This study aimed to determine the prevalence of brucellosis in cattle slaughtered in the Buea and Douala slaughterhouses and identify the Brucella species circulating among these animals. A total of 576 cattle serum samples were collected from the Buea and Douala slaughterhouses and analysed by ELISA. Following the ELISA assay, all samples were subjected to polymerase chain reaction (PCR) analysis. The bcsp31 gene primers were used for the genus-specific PCR. All bcsp31-positive samples were subjected to species-specific PCR. Primers targeting the IS711 gene sequence were used to identify the abortus-melitensis-ovis-suis species. The prevalence of brucellosis in both locations was 3.1% and 5.4% using the ELISA and PCR assays, respectively. Out of the 18 ELISA-positive samples, 5 (27%) were positive with PCR, while 26 (4.7%) of the ELISA-negative samples were positive with PCR. The Brucella species in circulation were identified using the Sanger sequencing technique. The sequences were 99.3% to 100% identical to the B. abortus strain BJ1-23 and the B. abortus strain BJ1-1 of the 31 kDa antigen (bcsp31) gene from India. This is the first report on the genotypic characterisation of Brucella species in Cameroon and confirms brucellosis in cattle at the Buea and Douala slaughterhouses.

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Genome-wide unique insertion sequences among five Brucella species and demonstration of differential identification of Brucella by multiplex PCR assay

Cited by 11 publications

References 27 publications

Rapid, ultrasensitive, and highly specific identification of Brucella abortus utilizing multiple cross displacement amplification combined with a gold nanoparticles-based lateral flow biosensor

Rapid, ultrasensitive, and highly specific identification of Brucella abortus utilizing multiple cross displacement amplification combined with a gold nanoparticles-based lateral flow biosensor

A Review on Brucellosis in Cameroon: Diagnostic Approaches, Epidemiology and Risk Factors for Infection

Identification and Prevalence of Brucella Species Circulating among Cattle Slaughtered in the Douala and Buea Municipalities of Cameroon

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