Rapid Multiplex PCR Assay for the Simultaneous Detection of the Brucella Genus, B. abortus, B. melitensis, and B. suis

Kumar, Sanjay; Tuteja, Urmil; Sarika, Kumari; Singh, Dhirendra Kumar; Kumar, Ashok; Kumar, Om

doi:10.4014/jmb.1007.07051

Cited by 29 publications

(21 citation statements)

References 25 publications

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“…Also, finding denaturation temperature, denaturation duration, and the annealing temperature are important subjects which are valuable and time consuming and require expert personnel. In contrast with other studies (20, 21), our study showed difference in sensitivity because of protocol conditions such as concentrations of primers, dNTPs, MgCl 2 , and Taq polymers. In the study by Da Costa et al , the specificity of primers on the 98 non- Brucella bacteria, mentioned that all organisms were negative for amplification of the BCSP31 gene (22).…”

Section: Discussioncontrasting

confidence: 99%

Evaluation of Different Primers for Detection of Brucella by Using PCR Method

2016

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IntroductionBrucellosis is a worldwide zoonosis and a significant cause of loss of health in humans and animals. Traditionally, classic diagnosis is carried out by isolation of Brucella, which is time-consuming, technically challenging and potentially dangerous. The aim of this study was to expand a molecular test that would be used for the develop detection of Brucella in a single reaction with high sensitivity and specificity, by targeting IS711element.MethodsThis study was carried out from 2015 to 2016 at the Ayatolla Rohani hospital in Babol, Iran. The present study was designed to develop PCR assay, based on IS711 gene for rapid diagnosis of Brucella spp. and immediate detection of Brucella, with high sensitivity and specificity. Four pairs of oligo-nucleotide primers with sizes of 547, 403, 291 and 127bp respectively, were planned to exclusively amplify the targeted genes of Brucella species.ResultsOur results show that, five PCR primers set up, would be helpful in amplifying the DNAs from the genus Brucella with high specificity and sensitivity so it can be 12 fg, for Brucella species to provide a valuable tool for diagnosis.ConclusionThis method can be more useful than serological and biochemical tests and in addition, this reduces the number of required tests more rapidly and economically.

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Section: Discussioncontrasting

confidence: 99%

Evaluation of Different Primers for Detection of Brucella by Using PCR Method

2016

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“…Rapid latex agglutination tests can also be useful in areas with limited laboratory capacity, and one study using culture-confirmed cases and negative controls demonstrated a sensitivity of 89% and a specificity of 98% [89]. PCR was effectively employed to rapidly detect Brucella DNA in the blood of six suspected cases which all subsequently met confirmed case definitions [90], and multiplex assays can expedite the confirmation and speciation of Brucella isolated by culture [91,92]. A real-time PCR assay that rapidly and accurately distinguishes Brucella from M. tuberculosis in body fluid and tissue specimens holds promise for clinical use [93].…”

Section: Clinical Diagnosis and Diagnostic Advancesmentioning

confidence: 99%

Brucellosis in low-income and middle-income countries

Rubach

Halliday

Cleaveland

et al. 2013

Current Opinion in Infectious Diseases

209

176

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Purpose of review Human brucellosis is a neglected, underrecognized infection of widespread geographic distribution. It causes acute febrile illness and a potentially debilitating chronic infection in humans, and livestock infection has substantial socioeconomic impact. This review describes new information regarding the epidemiology of brucellosis in the developing world and advances in diagnosis and treatment. Recent findings The highest recorded incidence of human brucellosis occurs in the Middle East and Central Asia. Fever etiology studies demonstrate brucellosis as a cause of undifferentiated febrile illness in the developing world. Brucellosis is a rare cause of fever among returning travelers, but is more common among travelers returning from the Middle East and North Africa. Sensitive and specific rapid diagnostic tests appropriate for resource-limited settings have been validated. Randomized controlled trials demonstrate that optimal treatment for human brucellosis consists of doxycycline and an aminoglycoside. Decreasing the burden of human brucellosis requires control of animal brucellosis, but evidence to inform the design of control programs in the developing world is needed. Summary Brucellosis causes substantial morbidity in human and animal populations. While improvements in diagnostic options for resource-limited settings and stronger evidence for optimal therapy should enhance identification and treatment of human brucellosis, prevention of human disease through control in animals remains paramount.

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“…Brucella melitensis, C. sakazakii and L. monocytogenes have been identified by conventional methods and real-time PCR (RTi-PCR) methods (Germini et al 2009;Kumar et al 2011;Jun-Ichi et al 2012). Conventional methods do not provide the desired accuracy, precision and speed, and sufficient information for quantitative detection of microorganisms compared with RTi-PCR technology.…”

Section: Introductionmentioning

confidence: 99%

Development and application of a new multiplex real‐time PCR assay for simultaneous identification of Brucella melitensis, Cronobacter sakazakii and Listeria monocytogenes in raw milk and cheese

Tutar

Akıncı

Akyol

2018

Int J of Dairy Tech

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Brucella melitensis, Cronobacter sakazakii and Listeria monocytogenes are important foodborne pathogens in milk and milk products, which are responsible for a variety of diseases that pose serious hazards to public health and food safety. The objective of this study was to develop a novel multiplex RTi‐PCR for the detection of B. melitensis, C. sakazakii and L. monocytogenes and to characterise the potential risk of these pathogens in raw milk and cheese. The raw milk (n = 25) and cheese samples (n = 20) were analysed by multiplex RTi‐PCR assay and detected for quantification of the three pathogens. In this study, B. melitensis, C. sakazakii and L. monocytogenes were simultaneously identified using BMEII0466, mms operon and hly as target genes, respectively. The multiplex RTi‐PCR assay that was developed showed good sensitivity and selectivity for the pathogenic microorganisms (r2 = 0.986–0.997). Multiplex RTi‐PCR results showed that most of the samples were contaminated with the pathogens screened.

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Rapid Multiplex PCR Assay for the Simultaneous Detection of the Brucella Genus, B. abortus, B. melitensis, and B. suis

Cited by 29 publications

References 25 publications

Evaluation of Different Primers for Detection of Brucella by Using PCR Method

Evaluation of Different Primers for Detection of Brucella by Using PCR Method

Brucellosis in low-income and middle-income countries

Development and application of a new multiplex real‐time PCR assay for simultaneous identification of Brucella melitensis, Cronobacter sakazakii and Listeria monocytogenes in raw milk and cheese

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