Comparison of Real-Time PCR with Conventional PCR and Culture to Assess the Efficacy of a Live Attenuated Salmonella enterica Serovar Typhimurium Vaccine Against Salmonella enterica Serovar Enteritidis in Commercial Leghorn Chicks Vaccinated Under Field and Laboratory Conditions

Parker, W. Drew; Lungu, A. Bwalya; Berghaus, Roy D.; Sellers, Holly S.; Alvarado, I. R.; Hofacre, Charles L.

doi:10.1637/9561-100410-reg.1

Cited by 10 publications

(1 citation statement)

References 25 publications

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“…Previous studies have described these same findings not only for Leishmania (Pourmohammadi et al, 2010; Sterkers et al, 2010; Eroglu et al, 2014) but for other microorganisms such as Helicobacter pylori (de Bruijn and Barker, 1992), Plasmodium spp (Gama et al, 2007), Salmonella enterica (Parker et al, 2011) and viruses such as ZIKV (Francino et al, 2006) and DENV (Bai et al, 2008; Faye et al, 2013). For the ARR, we observed that when kDNA is used a Ct between 5 and 27 is observed, a much lower range than the one reported for the other molecular markers ( Figure 2 ).…”

Section: Discussionmentioning

confidence: 55%

Analytical Performance of Four Polymerase Chain Reaction (PCR) and Real Time PCR (qPCR) Assays for the Detection of Six Leishmania Species DNA in Colombia

et al. 2017

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Leishmaniasis comprises a spectrum of parasitic diseases caused by protozoans of the genus Leishmania. Molecular tools have been widely employed for the detection of Leishmania due to its high sensitivity and specificity. However, the analytical performance of molecular platforms as PCR and real time PCR (qPCR) including a wide variety of molecular markers has never been evaluated. Herein, the aim was to evaluate the analytical performance of 4 PCR-based assays (designed on four different targets) and applied on conventional and real-time PCR platforms. We evaluated the analytical performance of conventional PCR and real time PCR, determining exclusivity and inclusivity, Anticipated Reportable Range (ARR), limit of detection (LoD) and accuracy using primers directed to kDNA, HSP70, 18S and ITS-1 targets. We observed that the kDNA was the most sensitive but does not meet the criterion of exclusivity. The HSP70 presented a higher LoD in conventional PCR and qPCR in comparison with the other markers (1 × 101 and 1 × 10-1 equivalent parasites/mL respectively) and had a higher coefficient of variation in qPCR. No statistically significant differences were found between the days of the test with the four molecular markers. The present study revealed that the 18S marker presented the best performance in terms of analytical sensitivity and specificity for the qPCR in the species tested (species circulating in Colombia). Therefore, we recommend to explore the analytical and diagnostic performance in future studies using a broader number of species across America.

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