A novel real-time PCR assay for specific detection of Brucella melitensis

Kaden, René; Ferrari, Sevinc; Alm, Erik; Wahab, Tara

doi:10.1186/s12879-017-2327-7

Cited by 40 publications

(31 citation statements)

References 23 publications

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“…; Kaden et al . ). Loop‐mediated isothermal amplification (LAMP) and real‐time PCR assays are sensitive molecular techniques for the detection of pathogen‐specific DNA from clinical specimens.…”

Section: Introductionmentioning

confidence: 97%

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Diagnostic utility of in‐house loop‐mediated isothermal amplification and real‐timePCRtargetingvirBgene for direct detection ofBrucella melitensisfrom clinical specimens

et al. 2019

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Aims In this present study, the utility of a newly developed loop‐mediated isothermal amplification (LAMP) and real‐time PCR assays designed to amplify the virB gene region of Brucella melitensis was evaluated from human clinical specimens. Methods and Results Fifty‐four culture‐confirmed cases of brucellosis and 54 culture negative but clinically suspected cases of brucellosis were included in the study. Whole blood, serum and other nonblood specimens were collected and subjected to blood culture using automatic blood culture system, serological tests, LAMP assay and real‐time PCR. Overall sensitivities of LAMP and real‐time PCR assays were 67·5 and 68·3% respectively. For nonblood clinical specimens, we noticed a marked increase in the sensitivities of LAMP (88·9%) and real‐time PCR (100%) assays. Conclusions Performance of LAMP and real‐time PCR was not satisfactory for whole‐blood specimens because of the low abundance of bacteria or DNA. On the other hand, using nonblood specimens, both the assays showed higher sensitivity and specificity which makes them a good alternative for the rapid diagnosis of human brucellosis. Significance and Impact of the Study The developed LAMP and real‐time PCR assays are a specific and rapid diagnostic tool for direct and early detection of Brucella in clinical specimens.

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Section: Introductionmentioning

confidence: 97%

“…; Kaden et al . ). Type IV secretion system (T4SS) is an important virulence determinant responsible for intracellular survival and multiplication of all the pathogenic species of Brucella (Boschiroli et al .…”

Section: Introductionmentioning

confidence: 97%

Diagnostic utility of in‐house loop‐mediated isothermal amplification and real‐timePCRtargetingvirBgene for direct detection ofBrucella melitensisfrom clinical specimens

et al. 2019

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“…In addition, B. melitensis is known to be the main human pathogen related to human outbreaks worldwide (Kaden et al . ). Cronobacter sakazakii is an opportunistic pathogen belonging to the family Enterobacteriaceae.…”

Section: Introductionmentioning

confidence: 97%

“…Brucellosis is widespread globally, especially in Mediterranean countries; it is also known as Malta fever or Mediterranean fever. In addition, B. melitensis is known to be the main human pathogen related to human outbreaks worldwide (Kaden et al 2017). Cronobacter sakazakii is an opportunistic pathogen belonging to the family Enterobacte-riaceae.…”

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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“…PCR can be performed with whole blood, serum, and tissue. The sensitivity and specificity of PCR are reported to be higher than those of the culture (18,19). …”

Section: Diagnosismentioning

confidence: 99%

An old foe still with us: Troubling issues with brucellosis

Buzğan¹,

Hasanoğlu²

2017

Med J Islamic World Acad Sci

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Clinical featuresBrucellosis can be subclinical, acute, subacute, and chronic. It can present itself with nonspecific symptoms such as fever, night sweats, loss of appetite, weight loss, weakness, headache, and polyarthritis (2,5-7). Brucellosis should be considered as a differential diagnosis for fevers of unknown origin. Debilitating conditions such as arthralgia, myalgia, and back pain affects around half of the patients. The incubation period is 2-3 weeks. Physical findings may vary depending on the duration of the disease. Systemic brucellosis sometimes can be complicated with meningitis, endocarditis, septic arthritis, and osteomyelitis. These complications are not extremely rare. A large meta-analysis reported the prevalence of endocarditis and neurobrucellosis as 1% and 4%, respectively (6). Osteoarticular complications are the most common complications of brucellosis (2,6,8).Relapse can occur 2-3 months after the treatment and can be detected with serologic tests and blood culture positivity. Relapse rate is common in cases treated with a single antibiotic regimen and short-term treatment. SUMMARYAlthough brucellosis is an old disease, it is still challenging for physicians. In this review, we discussed common pitfalls of brucellosis.

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A novel real-time PCR assay for specific detection of Brucella melitensis

Cited by 40 publications

References 23 publications

Diagnostic utility of in‐house loop‐mediated isothermal amplification and real‐timePCRtargetingvirBgene for direct detection ofBrucella melitensisfrom clinical specimens

Diagnostic utility of in‐house loop‐mediated isothermal amplification and real‐timePCRtargetingvirBgene for direct detection ofBrucella melitensisfrom clinical specimens

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

An old foe still with us: Troubling issues with brucellosis

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