A quantitative TaqMan PCR assay for the detection of Ureaplasma diversum

Marques, Lucas Miranda; Amorim, Aline Teixeira; Martins, Hellen Braga; Rezende, Izadora S.; Barbosa, Maysa Santos; Lobão, Tássia Neves; Campos, Guilherme Barreto; Timenetsky, Jorge

doi:10.1016/j.vetmic.2013.07.031

Cited by 16 publications

(12 citation statements)

References 15 publications

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“…The results presented here demonstrate that the bacterial load has been the major factor in the increased levels of pro-inflammatory mediators in the infection site. Higher bacterial load of U. diversum has been associated with more severe clinical conditions (Marques et al 2013). Infected animals respond with increased levels of TNF-α and IFN-γ production.…”

Section: Discussionmentioning

confidence: 99%

Intra-uterine experimental infection by Ureaplasma diversum induces TNF-α mediated womb inflammation in mice

Silva

Ferreira

Oliveira

et al. 2016

An. Acad. Bras. Ciênc.

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Ureaplasma diversum is an opportunistic pathogen associated with uterine infl ammation, impaired embryo implantation, infertility, abortions, premature birth of calves and neonatal pneumonia in cattle. It has been suggested that the intra-uterine infection by Ureaplasma diversum can cause vascular changes that hinder the success of pregnancy. Thus, the aim of this study was to evaluate the changes of intrauterine site of A/J mice in estrus or proestrus phase inoculated with Ureaplasma diversum. The infection was monitored at 24, 48 and 72 hours by the PCR methodology to detect the Ureaplasma in the inoculation site and the profi le of circulating blood cells. Morphological changes, intensity of infl ammation and the production of cytokines were compared. The infected mice showed local infl ammation through the production of IFN-γ and TNF-α. Ureaplasma diversum infections in the reproductive tract of studied mice seemed to be associated with the production of pro-infl ammatory cytokines in uterine parenchyma. The levels of TNF-α of infected mice were dependent on the bacterial load of inoculated Ureaplasma. Uterine experimental infections by Ureaplasma diversum have not been mentioned yet and herein we presented the fi rst report of an intrauterine infection model in mice.

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Section: Discussionmentioning

confidence: 99%

Intra-uterine experimental infection by Ureaplasma diversum induces TNF-α mediated womb inflammation in mice

Silva

Ferreira

Oliveira

et al. 2016

An. Acad. Bras. Ciênc.

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“…Each primer-probe set was designed to detect a single target pathogen. As shown in Table 1, seven primer-probe sets were adopted from a previously reported real-time PCR assay [2,18,20,21,26,32,33]. The remaining nine primer-probe sets were newly designed in this study.…”

Section: Primer and Probe Designmentioning

confidence: 99%

Development of a one-run real-time PCR detection system for pathogens associated with bovine respiratory disease complex

Kishimoto

Tsuchiaka

Rahpaya

et al. 2017

The Journal of Veterinary Medical Science

102

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Bovine respiratory disease complex (BRDC) is frequently found in cattle worldwide. The etiology of BRDC is complicated by infections with multiple pathogens, making identification of the causal pathogen difficult. Here, we developed a detection system by applying TaqMan real-time PCR (Dembo respiratory-PCR) to screen a broad range of microbes associated with BRDC in a single run. We selected 16 bovine respiratory pathogens (bovine viral diarrhea virus, bovine coronavirus, bovine parainfluenza virus 3, bovine respiratory syncytial virus, influenza D virus, bovine rhinitis A virus, bovine rhinitis B virus, bovine herpesvirus 1, bovine adenovirus 3, bovine adenovirus 7, Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, Trueperella pyogenes, Mycoplasma bovis and Ureaplasma diversum) as detection targets and designed novel specific primer-probe sets for nine of them. The assay performance was assessed using standard curves from synthesized DNA. In addition, the sensitivity of the assay was evaluated by spiking solutions extracted from nasal swabs that were negative by Dembo respiratory-PCR for nucleic acids of pathogens or synthesized DNA. All primer-probe sets showed high sensitivity. In this study, a total of 40 nasal swab samples from cattle on six farms were tested by Dembo respiratory-PCR. Dembo respiratory-PCR can be applied as a screening system with wide detection targets.

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“…Improved sensitivity in detecting Tritrichomonas fetus in smegma and vaginal mucous was demonstrated using PCR compared to culture. Quantitative PCR testing for Ureaplasma diversum was 100-fold more sensitive than conventional PCR (Marques et al, 2013). In addition, buccal swabs can also be collected to test for a range of inherited diseases including collie eye anomaly (Chang et al, 2010), hereditary cataract (Mellersh et al, 2006), and ceroid lipofuscinosis in dogs (Karli et al, 2014).…”

Section: Swabs From Mucosal Surfacesmentioning

confidence: 99%

Moving past serology: Diagnostic options without serum

Reichel

Lanyon

Hill³

2016

The Veterinary Journal

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Detecting antibodies formed in serum in response to infection is the traditional function of serology. Diagnostic modalities have included complement fixation tests, agar gel immune-diffusion, radioimmunoassay, ELISA and immunofluorescence. More recent technology now allows for the direct detection of pathogens by PCR. This review details the options for diagnostic testing using specimen types other than serum, identifying the advantages and disadvantages of these options and providing evidence for more widespread use of these techniques and specimen types.

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A quantitative TaqMan PCR assay for the detection of Ureaplasma diversum

Cited by 16 publications

References 15 publications

Intra-uterine experimental infection by Ureaplasma diversum induces TNF-α mediated womb inflammation in mice

Intra-uterine experimental infection by Ureaplasma diversum induces TNF-α mediated womb inflammation in mice

Development of a one-run real-time PCR detection system for pathogens associated with bovine respiratory disease complex

Moving past serology: Diagnostic options without serum

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