Differentiation of Brucella abortus and B. melitensis biovars using PCR-RFLP and REP-PCR

Amoupour, Moein; Nezamzadeh, Fatemeh; Bialvaei, Abed Zahedi; Sedighi, Mansour; Jazi, Faramarz Masjedian; Alikhani, Mohammad Yousef; Mirnejad, Reza

doi:10.1016/j.nmni.2019.100589

Cited by 5 publications

(7 citation statements)

References 22 publications

(28 reference statements)

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“…We attempted to evaluate the molecular markers reported by different authors; however, a contradiction or failure to reproduce them limits the applicability of these markers. The primers designed in the current study, along with IS711 ( 28 ), were able to diagnose brucellosis in 1.27% (CI: 0.56–3.11) of animal samples (1.05% samples were positive for at least one serological and molecular test). The designed primers successfully amplified in the positive control ( B. abortus DNA provided by IVRI, Bareilly), and the amplicons of unidentified samples sequenced using Sanger sequencing confirmed the applicability of these markers.…”

Section: Discussionmentioning

confidence: 74%

“…Primer specificity was analyzed for different species of Brucella and checked for any non-specific binding in the host genome (five livestock species sampled for the current study) using Primer blast ( 27 ). After optimization with unknown samples and positive control sample, we finally used the following primer sets in the present study IS711 primers ( 28 ) and genus-specific primer 1 (GSP-1; designed for the present study as mentioned earlier) ( Table 1 ). The DNA sequence comparisons with the GenBank database were searched and assessed for species or genus assignment using BLAST search ( 29 ).…”

Section: Methodsmentioning

confidence: 99%

“… a adopted from Amoupour et al ( 28 ); Ba, Brucella abortus ; Bm, B. melitensis ; GSP, genus-specific primer; GSP-1 was designed from a conserved region of the reference sequence of Brucella abortus (NC_007624.1). …”

Section: Methodsmentioning

confidence: 99%

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Serological and molecular prevalence of Brucella spp. among livestock species in Rajasthan, India

Meena¹,

Sharma²,

Bishnoi³

et al. 2023

Front. Vet. Sci.

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A seroprevalence and molecular study was carried out in six districts of the state of Rajasthan, India to detect brucellosis in major livestock species. This study involves the testing of 3,245 livestock samples using the Rose Bengal Plate Test (RBPT), Indirect Enzyme-Linked Immunosorbent Assay (i-ELISA), and genus-specific polymerase chain reaction (PCR) markers for molecular diagnosis of the disease. In the tested samples, seroprevalence was 5.06% (CI: 1.96–8.15) using the RBPT test and 6.88% (CI: 1.98–11.78) using the i-ELISA test, while the cumulative seroprevalence (RBPT and i-ELISA) was 3.63% (CI: 0.44–6.83). The prevalence of the disease was 1.27% (CI: 0.56–3.11) when tested using molecular markers. The highest prevalence of brucellosis was detected in Cattle (7.00, 3.22%), followed by camels (5.50, 2.50%), buffalo (2.66, 0.00%), sheep (2.43, 0.41%), and goats (0.58, 0.23%) when serological (cumulative) and molecular diagnosis were considered preferred methods of detection. Cattle (3.22%) and camels (2.50%) also showed a high prevalence of disease when tested using molecular markers. The results of this study reveal that cattle, camel, and sheep brucellosis is prevalent in the study areas.

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Section: Discussionmentioning

confidence: 74%

Section: Methodsmentioning

confidence: 99%

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Serological and molecular prevalence of Brucella spp. among livestock species in Rajasthan, India

Meena¹,

Sharma²,

Bishnoi³

et al. 2023

Front. Vet. Sci.

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“…Rep-PCR is a conventional and broadly used technique in microbiology and genetic diversity. Such as plant pathogenic bacteria wilt of Solanaceaous vegetables R. solanacearum (Hossain et al 2022), fungal disease strawberry anthracnose (Karimi et al 2019), and zoonotic disease brucellosis (Amoupour et al 2019). In this study, using rep-PCR technique, the genetic diversity of 60 walnut bacterial-associated bacteria from different regions was investigated through three sets of primers (ERIC, BOX, and REP).…”

Section: Discussionmentioning

confidence: 99%

Short Communication: Genetic diversity of walnut blight-associated bacteria from China using rep-PCR

et al. 2022

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Abstract. Zheng C, Zhu J, Yang X, Fu B. 2022. Short Communication: Genetic diversity of walnut blight-associated bacteria from China using rep-PCR. Biodiversitas 23: 5681-5686. Walnut blight is caused by the bacterium Xanthomonas arboricola pv. juglandis (Xaj), is one of the most serious diseases in walnut production around the world. Understanding the genetic diversity of walnut blight pathogen is of great significance to developing a scientific and reasonable prevention and control strategy. The rep-PCR (Repetitive Sequence Polymerase Chain Reaction) technique is a powerful tool for analyzing plant pathogens. In this paper, using three pairs of rep-PCR universal primers (ERIC, BOX, and REP), a genetic diversity analysis was carried out on walnut blight-associated bacteria mainly isolated from different counties of Hubei Province, China. A study of 60 walnut blight-associated bacteria isolated in 2016 found that they had a high genetic similarity among them. There is no clear correlation between the genetic diversity of these isolates and their geographical or tissue origins. Interestingly, these clusters can also distinguish copper-resistant isolates from sensitive ones. Therefore, rep-PCR is a powerful tool to study the population of walnut pathogenic bacteria.

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“…However, these procedures are expensive and are not readily available, particularly in developing countries such as Egypt. Alternatively, several studies have employed more cost effective discrimination approaches, such as IS 711 gene sequence variations, PCR restriction fragment length polymorphism (PCR-RFLP), and Repetitive element sequence-based PCR (rep-PCR) [ [10] , [11] , [12] ]. However, the high genetic similarity between Brucella species can occasionally reduce the efficiency of these tools in discriminating between isolates of same species, especially when these tools were used individually [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

Risk factors and Molecular genotyping of Brucella melitensis strains recovered from humans and their owned cattle in Upper Egypt

et al. 2021

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Brucellosis is a zoonosis that has a devastating impact on the economy and public health, particularly in the Middle East, including Egypt. This study aimed to define risk factors associated with brucellosis in humans and in their cattle in Fayoum governorate - Upper Egypt. Also, molecular genotyping of recovered Brucella isolates from human cases and their cattle to assess the potential cross-species transmission in the study region. Data were obtained via double matched case–control studies for brucellosis in humans (106 cases and 160 controls) and in their cattle (78 cattle cases and 105 cattle controls). The results of multivariate regression analysis revealed that predictors of human brucellosis were animal-related occupations (OR 2.1, P 0.02), previous infection in other household members (OR 3.2, P 0.007), eating home-made soft cheese (OR 2.3, P 0.03), and exposure to cattle abortions (OR 6.9, P < 0.001). For cattle, predictors of brucellosis were maturity ≥2 years of age (OR 2.9, P 0.01), ≥2 animals reared by the same household (OR 3.7–6.9, P ≤ 0.001), and recent abortion (OR 15.2, P 0.01). Twelve Brucella isolates were recovered from eight human cases (7.5%, 8/106) and four cattle cases (6.2%, 4/65). All isolates were B. melitensis biovar 3. Analysis of the IS711 gene sequence revealed complete homology (100%) between isolates. Six virulence genes were utilized for virutyping: virB (100%), omp25 (100%), amiC (100%), ure (91.7%), wbkA (91.7%), and bvfA (75%). Virutyping revealed four virutypes: V1 (lack bvfA , 16.7%), V2 (harbored all genes, 66.7%), V3 (lack wbkA , 8.3%), and V4 (lack wbkA and ure , 8.3%). Repetitive extragenic palindromic PCR (REP-PCR) typing revealed two REP types. Combined REP-PCR/virulence genotyping revealed five different genotypes (G1–G5) for the detected isolates and a unique genotype for the reference strain (G6, B. melitensis bv3 Ether). Human and cattle isolates from the same household had matched genotypes. In conclusion, there were widespread risk factors among the cases studied. Health education for high-risk groups is essential for disease prevention, and combined REP-PCR/virulence genotyping is a quick tool for traceability, particularly in developing countries endemic with brucellosis as Egypt.

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Differentiation of Brucella abortus and B. melitensis biovars using PCR-RFLP and REP-PCR

Cited by 5 publications

References 22 publications

Serological and molecular prevalence of Brucella spp. among livestock species in Rajasthan, India

Serological and molecular prevalence of Brucella spp. among livestock species in Rajasthan, India

Short Communication: Genetic diversity of walnut blight-associated bacteria from China using rep-PCR

Risk factors and Molecular genotyping of Brucella melitensis strains recovered from humans and their owned cattle in Upper Egypt

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