One-step multiplex RT-qPCR assay for the detection of Peste des petits ruminants virus, Capripoxvirus, Pasteurella multocida and Mycoplasma capricolum subspecies (ssp.) capripneumoniae

Settypalli Tirumala Bharani Kumar, Lamien Charles Euloge, Spergser Joachim, Lelenta Mamadou, Wade Abel, Gelaye Esayas, Loitsch Angelika, Minoungou Germaine, Thiaucourt François, Diallo Adama. 2016. One-step multiplex RT-qPCR assay for the detection of Peste des petits ruminants virus, Capripoxvirus, Pasteurella multocida and Mycoplasma capricolum subspecies (ssp.) capripneumoniae. PloS One, 11 (4):e0153688, 14 p.

Quartile : Q1, Sujet : MULTIDISCIPLINARY SCIENCES

Liste HCERES des revues (en SHS) : oui

Thème(s) HCERES des revues (en SHS) : Psychologie-éthologie-ergonomie; Staps

Résumé : Respiratory infections, although showing common clinical symptoms like pneumonia, are caused by bacterial, viral or parasitic agents. These are often reported in sheep and goats populations and cause huge economic losses to the animal owners in developing countries. Detection of these diseases is routinely done using ELISA or microbiological methods which are being reinforced or replaced by molecular based detection methods including multiplex assays, where detection of different pathogens is carried out in a single reaction. In the present study, a one-step multiplex RT-qPCR assay was developed for simultaneous detection of Capripoxvirus (CaPV), Peste de petits ruminants virus (PPRV), Pasteurella multocida (PM) and Mycoplasma capricolum ssp. capripneumonia (Mccp) in pathological samples collected from small ruminants with respiratory disease symptoms. The test performed efficiently without any cross-amplification. The multiplex PCR efficiency was 98.31%, 95.48%, 102.77% and 91.46% whereas the singleplex efficiency was 93.43%, 98.82%, 102.55% and 92.0% for CaPV, PPRV, PM and Mccp, respectively. The correlation coefficient was greater than 0.99 for all the targets in both multiplex and singleplex. Based on cycle threshold values, intra and inter assay variability, ranged between the limits of 2%–4%, except for lower concentrations of Mccp. The detection limits at 95% confidence interval (CI) were 12, 163, 13 and 23 copies/reaction for CaPV, PPRV, PM and Mccp, respectively. The multiplex assay was able to detect CaPVs from all genotypes, PPRV from the four lineages, PM and Mccp without amplifying the other subspecies of mycoplasmas. The discriminating power of the assay was proven by accurate detection of the targeted pathogen (s) by screening 58 viral and bacterial isolates representing all four targeted pathogens. Furthermore, by screening 81 pathological samples collected from small ruminants showing respiratory disease symptoms, CaPV was detected in 17 samples, PPRV in 45, and PM in six samples. In addition, three samples showed a co-infection of PPRV and PM. Overall, the one-step multiplex RT-qPCR assay developed will be a valuable tool for rapid detection of individual and co-infections of the targeted pathogens with high specificity and sensitivity.

Mots-clés Agrovoc : peste des petits ruminants, Mycoplasma capricolum, capripoxvirus, Pasteurella multocida

Classification Agris : L73 - Maladies des animaux

Champ stratégique Cirad : Axe 4 (2014-2018) - Santé des animaux et des plantes

Auteurs et affiliations

• Settypalli Tirumala Bharani Kumar, IAEA (AUT)
• Lamien Charles Euloge, IAEA (AUT)
• Spergser Joachim, University of Veterinary Medicine Vienna (AUT)
• Lelenta Mamadou, IAEA (AUT)
• Wade Abel, LANAVET (CMR)
• Gelaye Esayas, IAEA (AUT)
• Loitsch Angelika, Institute for Veterinary Disease Control (AUT)
• Minoungou Germaine, Laboratoire National d'Elevage (BFA)
• Thiaucourt François, CIRAD-BIOS-UMR CMAEE (FRA)
• Diallo Adama, CIRAD-BIOS-UMR CMAEE (SEN)

Source : Cirad-Agritrop (https://agritrop.cirad.fr/581042/)

Métriques

[ Page générée et mise en cache le 2024-04-03 ]