Detection of all Chlamydophila and Chlamydia spp. of veterinary interest using species-specific real-time PCR assays

Pantchev, Alexandra; Sting, Reinhard; Bauerfeind, Rolf; Tyczka, Judith; Sachse, Konrad

doi:10.1016/j.cimid.2009.08.002

Cited by 150 publications

(139 citation statements)

References 26 publications

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“…The analytical specificity of the C. abortus and C. pecorum species-specific qPCR 117 assays has been described previously (Pantchev et al, 2009(Pantchev et al, , 2010, but was further assessed 118 by testing DNA from a wide range of bacterial and parasitic species. To determine the 119 sensitivity of the assay, 10-fold serial dilutions of plasmids were prepared containing the 120 cloned PCR products amplified from C. abortus or C. pecorum, these were spiked into faecal 121 samples and the DNA was extracted and amplified as described above.…”

Section: Specificity and Sensitivity 116mentioning

confidence: 99%

Longitudinal prevalence and faecal shedding of Chlamydia pecorum in sheep

Yang

Jacobson

Gardner

et al. 2014

The Veterinary Journal

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20The prevalence and faecal shedding of Chlamydia spp. in sheep in Australia has not 21 been well described. Two species-specific quantitative PCRs (qPCR) targeting the chlamydial 22 outer membrane protein cell surface antigen gene (ompA) were validated and used to 23 determine the prevalence and faecal shedding of C. abortus and C. pecorum from faecal 24 samples of lambs at three sampling times (weaning, post-weaning and pre-slaughter) from 25

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Section: Specificity and Sensitivity 116mentioning

confidence: 99%

Longitudinal prevalence and faecal shedding of Chlamydia pecorum in sheep

Yang

Jacobson

Gardner

et al. 2014

The Veterinary Journal

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“…The negative samples were treated as described earlier and inoculated into chicken egg embryos for another passage. 20 The positive PCR products of the nmPCR assay were cloned into pMD19-T vector. Positive clones were sequenced and multi-aligned with the appropriate reference sequences (C. suis AF269274; Ch.…”

Section: Pathogen Isolation and Sequencingmentioning

confidence: 99%

“…of veterinary importance needs 6 separate, species-specific real-time PCR assays. 20 For the diagnosis of enzootic abortion, a multiplex PCR for the differential detection of Ch. abortus and Chlamydophila pecorum and a duplex real-time PCR for the specific detection of Chlamydophila psittaci and Ch.…”

Section: Introductionmentioning

confidence: 99%

A nested multiplex polymerase chain reaction assay for the differential identification of three zooanthroponotic chlamydial strains in porcine swab samples

Wang

Nie

et al. 2011

J VET Diagn Invest

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Abstract. Porcine chlamydial infection is an enzootic infectious disease caused by multiple members of the family Chlamydiaceae (e.g. Chlamydophila abortus, Chlamydia suis, and Chlamydophila pneumoniae). Rapid and accurate differentiation of these pathogens is critical in the control and prevention of disease. The aim of the current study was to develop a nested multiplex polymerase chain reaction (nmPCR) assay to simultaneously detect the 3 chlamydial pathogens in clinical samples. In the first round of the nmPCR, 1 pair of family-specific primers were used to amplify the 1,100 base pair (bp) fragment of chlamydial ompA gene. In the second round of the nmPCR, 4 inner primers were designed for Ch. abortus, C. suis, and Ch. pneumoniae. Each pathogen produced a specific amplicon with a size of 340 bp, 526 bp, and 267 bp respectively. The assay was sensitive and specific for detecting target pathogens in both cell cultures and clinical specimens. The results, incorporated with the improved rapid DNA extraction protocol, suggest that the nmPCR could be a promising assay for differential identification of different chlamydial strains in pigs.

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“…For rapid and specific diagnosis, direct detection by polymerase chain reaction (PCR) has been proven to be an important tool. 5,11,14 To better use diagnostic resources, the aim of the current study was to develop a multiplex real-time PCR for the simultaneous and rapid detection of Chlamydia spp., C. burnetii, and N. caninum.…”

mentioning

confidence: 99%

“…14 and N. caninum were used. 5 For C. burnetii, a previously described assay that targets the iso citrate dehydrogenase gene (icd ) 11 was used.…”

mentioning

confidence: 99%

Simultaneous detection of Chlamydia spp., Coxiella burnetii, and Neospora caninum in abortion material of ruminants by multiplex real-time polymerase chain reaction

2013

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Chlamydia spp., Coxiella burnetii, and Neospora caninum are responsible for reproductive diseases and are closely linked with high abortion rates in ruminants. Furthermore, C. burnetii and Chlamydia spp. have zoonotic potential. A real-time polymerase chain reaction (PCR) assay was developed for the simultaneous detection of Chlamydia spp., C. burnetii, and N. caninum. The detection of beta-actin as internal control in the same PCR reaction provides additional information about sample quality by detecting the presence of PCR inhibitors. The multiplex real-time PCR developed in the current study shows a greater sensitivity compared to previously used single-target PCR reactions with a reproducible detection limit of 0.13 plasmid copies per PCR for each target. Additional parallel amplification of all detectable pathogens did not adversely impact sensitivity. This new multiplex PCR allows the highly sensitive, cost-effective, and rapid detection of 3 important pathogens and has the potential to be a useful time-saving tool in the routine diagnosis of abortion cases in ruminants.

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Detection of all Chlamydophila and Chlamydia spp. of veterinary interest using species-specific real-time PCR assays

Cited by 150 publications

References 26 publications

Longitudinal prevalence and faecal shedding of Chlamydia pecorum in sheep

Longitudinal prevalence and faecal shedding of Chlamydia pecorum in sheep

A nested multiplex polymerase chain reaction assay for the differential identification of three zooanthroponotic chlamydial strains in porcine swab samples

Simultaneous detection of Chlamydia spp., Coxiella burnetii, and Neospora caninum in abortion material of ruminants by multiplex real-time polymerase chain reaction

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