Interlaboratory Comparison of Real-Time Polymerase Chain Reaction Methods to Detect <i>Coxiella Burnetii</i>, The Causative Agent of Q Fever

Jones, Rosemary; Hertwig, Stefan; Pitman, James; Vipond, Richard; Aspán, Anna; Bölske, Göran; McCaughey, Conall; McKenna, James; Rotterdam, Bart J. van; Bruin, Arnout de; Ruuls, Robin; Buijs, Rob; Roest, Hendrik Jan; Sawyer, Jason

doi:10.1177/104063871102300118

Cited by 20 publications

(13 citation statements)

References 23 publications

(42 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Detection of genetic material by PCR does not always imply viable infective bacteria ( 6 ). However, because the infectious dose of C. burnetii is low ( 10 ), our findings support the use of preventive hygiene measures ( 4 ) to minimize zoonotic risk when handling roe deer. The 2 MLVA-typed strains provided no evidence for spillover of the predominant strain involved in the Q fever epidemic in the Netherlands.…”

supporting

confidence: 57%

Coxiella burnetiiInfection in Roe Deer during Q Fever Epidemic, the Netherlands

Rijks¹,

Roest²,

Tulden³

et al. 2011

Emerg. Infect. Dis.

View full text Add to dashboard Cite

supporting

confidence: 57%

Coxiella burnetiiInfection in Roe Deer during Q Fever Epidemic, the Netherlands

Rijks¹,

Roest²,

Tulden³

et al. 2011

Emerg. Infect. Dis.

View full text Add to dashboard Cite

“…In addition, the multiplex qPCR assay was tested in a ring trial for C. burnetii DNA detection. Ten DNA samples, obtained from fetal fluids and placenta materials of ovine, caprine, and bovine origin, were compared between seven laboratories (14).…”

Section: Resultsmentioning

confidence: 99%

“…DNA of C. burnetii strain Nine Mile RSA phase I and isolates from mouse spleen (EP3, Russia, 1958, Apodemus flavicollis), and tick (EP5, Slovakia, 1968, Dermacentor marginatus) were used in experiments to test the performance of the qPCR assay. In addition, the developed multiplex qPCR assay was tested in a ring trial for C. burnetii DNA detection, in which 10 DNA samples from various origins were compared among seven laboratories (14).…”

Section: Methodsmentioning

confidence: 99%

Detection of Coxiella burnetii in Complex Matrices by Using Multiplex Quantitative PCR during a Major Q Fever Outbreak in The Netherlands

Bruin

Groot

Heer

et al. 2011

Appl Environ Microbiol

View full text Add to dashboard Cite

Q fever, caused by Coxiella burnetii, is a zoonosis with a worldwide distribution. A large rural area in the southeast of the Netherlands was heavily affected by Q fever between 2007 and 2009. This initiated the development of a robust and internally controlled multiplex quantitative PCR (qPCR) assay for the detection of C. burnetii DNA in veterinary and environmental matrices on suspected Q fever-affected farms. The qPCR detects three C. burnetii targets (icd, com1, and IS1111) and one Bacillus thuringiensis internal control target (cry1b). Bacillus thuringiensis spores were added to samples to control both DNA extraction and PCR amplification. The performance of the qPCR assay was investigated and showed a high efficiency; a limit of detection of 13.0, 10.6, and 10.4 copies per reaction for the targets icd, com1, and IS1111, respectively; and no crossreactivity with the nontarget organisms tested. Screening for C. burnetii DNA on 29 suspected Q fever-affected farms during the Q fever epidemic in 2008 showed that swabs from dust-accumulating surfaces contained higher levels of C. burnetii DNA than vaginal swabs from goats or sheep. PCR inhibition by coextracted substances was observed in some environmental samples, and 10-or 100-fold dilutions of samples were sufficient to obtain interpretable signals for both the C. burnetii targets and the internal control. The inclusion of an internal control target and three C. burnetii targets in one multiplex qPCR assay showed that complex veterinary and environmental matrices can be screened reliably for the presence of C. burnetii DNA during an outbreak.

show abstract

“…For the regional bulk milk survey of dairy herds on the Isle of Gotland, the national goat survey as well as the vaginal swab survey in sheep, an in-house real-time PCR protocol was used (Swedish Institute for Communicable Disease Control; Talar Boskani, personal communication). The two PCR protocols performed equally well when evaluated in an interlaboratory comparison of real-time PCR methods to detect C. burnetii [16]. …”

Section: Methodsmentioning

confidence: 99%

Surveys on Coxiella burnetii infections in Swedish cattle, sheep, goats and moose

et al. 2014

Self Cite

View full text Add to dashboard Cite

BackgroundQ fever is a zoonotic disease caused by the bacterium Coxiella burnetii. Prevalence data in ruminant species are important to support risk assessments regarding public and animal health. The aim was to investigate the presence of or exposure to C. burnetii in cattle, sheep, goats and moose, and to compare two enzyme-linked immunosorbent assays (ELISAs). National surveys of antibodies against C. burnetii were performed for dairy cattle (n=1537), dairy goats (n=58) and sheep (n=518). Bovine samples consisted of bulk milk, caprine of pooled milk, and ovine of pooled serum. Antibodies were investigated in moose samples (n=99) from three regions. A one-year regional cattle bulk milk survey was performed on the Isle of Gotland (n=119, four occasions). Cattle, sheep and goat samples were analysed with indirect ELISA and moose samples with complement fixation test. For the sheep, goat, and parts of the cattle survey, samples were run in parallel by ELISAs based on antigens from infected ruminants and ticks. Bulk milk samples from the regional cattle survey and vaginal swabs from a subset of the sheep herds (n=80) were analysed for the agent by polymerase chain reaction. Spatial clustering was investigated in the national cattle survey.ResultsThe prevalence of antibodies in dairy herds was 8.2% with large regional differences. High risk clusters were identified in the southern regions. The prevalence among dairy herds on the Isle of Gotland varied from 55.9% to 64.6% and 46.4% to 58.9.0% for antibodies and agent, respectively, overall agreement between agent and antibodies was 85.2%. The prevalence of antibodies in sheep was 0.6%, the agent was not detected the vaginal swabs. Antibodies were not detected in goats or moose, although parts of the moose samples were collected in an area with high prevalence in cattle. The overall agreement between the two ELISAs was 90.4%.ConclusionsThe prevalence of antibodies against C. burnetii in dairy cattle in Sweden shows large regional differences. The results suggest that C. burnetii is a rare pathogen among Swedish moose, dairy goat and sheep. ELISAs based on ruminant and tick antigen performed in a similar manner under Swedish conditions.

show abstract

Interlaboratory Comparison of Real-Time Polymerase Chain Reaction Methods to Detect Coxiella Burnetii, The Causative Agent of Q Fever

Cited by 20 publications

References 23 publications

Coxiella burnetiiInfection in Roe Deer during Q Fever Epidemic, the Netherlands

Coxiella burnetiiInfection in Roe Deer during Q Fever Epidemic, the Netherlands

Detection of Coxiella burnetii in Complex Matrices by Using Multiplex Quantitative PCR during a Major Q Fever Outbreak in The Netherlands

Surveys on Coxiella burnetii infections in Swedish cattle, sheep, goats and moose

Contact Info

Product

Resources

About