scholarly journals MLST typing of Pasteurella multocida associated with haemorrhagic septicaemia and development of a real-time PCR specific for haemorrhagic septicaemia associated isolates

2014 ◽  
Vol 170 (3-4) ◽  
pp. 335-341 ◽  
Author(s):  
Andreas Petersen ◽  
Magne Bisgaard ◽  
Kirsty Townsend ◽  
Henrik Christensen
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pasteurella Multocida ◽  
Haemorrhagic Septicaemia

scholarly journals Estimating Clinically Relevant Cut-Off Values for a High-Throughput Quantitative Real-Time PCR Detecting Bacterial Respiratory Pathogens in Cattle

2021 ◽  
Vol 8 ◽  
Author(s):  
Alicia F. Klompmaker ◽  
Maria Brydensholt ◽  
Anne Marie Michelsen ◽  
Matthew J. Denwood ◽  
Carsten T. Kirkeby ◽  
...  
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Real Time Pcr ◽  
Pasteurella Multocida ◽  
Age Groups ◽  
Therapeutic Interventions ◽  
Respiratory Pathogens ◽  
Logistic Regression Models ◽  
Histophilus Somni ◽  
Nasal Swabs

Bovine respiratory disease (BRD) results from interactions between pathogens, environmental stressors, and host factors. Obtaining a diagnosis of the causal pathogens is challenging but the use of high-throughput real-time PCR (rtPCR) may help target preventive and therapeutic interventions. The aim of this study was to improve the interpretation of rtPCR results by analysing their associations with clinical observations. The objective was to develop and illustrate a field-data driven statistical method to guide the selection of relevant quantification cycle cut-off values for pathogens associated with BRD for the high-throughput rtPCR system “Fluidigm BioMark HD” based on nasal swabs from calves. We used data from 36 herds enrolled in a Danish field study where 340 calves within pre-determined age-groups were subject to clinical examination and nasal swabs up to four times. The samples were analysed with the rtPCR system. Each of the 1,025 observation units were classified as sick with BRD or healthy, based on clinical scores. The optimal rtPCR results to predict BRD were investigated for Pasteurella multocida, Mycoplasma bovis, Histophilus somni, Mannheimia haemolytica, and Trueperella pyogenes by interpreting scatterplots and results of mixed effects logistic regression models. The clinically relevant rtPCR cut-off suggested for P. multocida and M. bovis was ≤ 21.3. For H. somni it was ≤ 17.4, while no cut-off could be determined for M. haemolytica and T. pyogenes. The demonstrated approach can provide objective support in the choice of clinically relevant cut-offs. However, for robust performance of the regression model sufficient amounts of suitable data are required.

scholarly journals Detection of Colistin Resistance in Mannheimia haemolytica & Pasteurella multocida Isolates from Ruminants in Morocco

2021 ◽  
Vol 41 (01) ◽  
pp. 127-131
Author(s):  
G Sebbar
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pasteurella Multocida ◽  
Mannheimia Haemolytica ◽  
Gram Negative Bacteria ◽  
Colistin Resistance ◽  
Gram Negative ◽  
Pcr Screening ◽  
Antimicrobial Sensitivity ◽  
Resistance Rates

Colistin is an important drug of last resort against lethal infections with multidrugresistant gram-negative bacteria, this antibiotic interacts on phosphate groups of lipopolysaccharides present on the outer membrane of gram-negative bacteria. This first mechanism of transferable colistin resistance involves a gene called mcr-1. Here, we described different tools such as Colistin susceptibility test, Minimal Inhibitory Concentration to find the colistin profiles of Mannheimia haemolytica and Pasteurella multocida serogroup A Moroccan strain isolated from nasal swabs and lung taken from sheep, goat and cattle with respiratory diseases during January 2015 to December 2017 in six different regions, in addition, we investigated either real time PCR to detect mcr-1 gene. Antimicrobial sensitivity test, was achieved for Fortyone isolates, the resistance rates of isolates from Pasteurellaceae species were between 59 and 71%, which respectively correspond to Mannheimia haemolytica and Pasteurella multocida segroup A. The sensitive ratios were between 29 and 41% which respectively correspond to Pasteurella multocida segroup A and Mannheimia haemolytica. Also, the MIC test was done against colistin, the results showed a resistant profile with a MIC >=64 μg/ml except for 3 strains from ruminants which have a value <2 μg/ml. The real time PCR screening test was detected with ct values ranging from 23 to 31 and confirmed the results obtained by MIC test. To our knowledge the present study is the first study which reports the resistance to colistin in Pasteurellaceae species strains isolated from ruminants in Morocco, our results suggest the necessity and the urgency for establishing a national program for monitoring antibacterial resistance against colistin

Evaluation of a SYBR Green Real-Time PCR Assay for Specific Detection of Pasteurella multocida in Culture and Tissue Samples from Sheep

2020 ◽  
Author(s):  
Jyoti Kumar ◽  
G. G. Sonawane ◽  
Fateh Singh ◽  
S. Jegaveera Pandian ◽  
Rajiv Kumar
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pasteurella Multocida ◽  
Bacterial Species ◽  
Sybr Green ◽  
Economic Losses ◽  
Conventional Pcr ◽  
Specific Detection ◽  
Pcr Assay ◽  
Tissue Samples

Pasteurella multocida is one of the bacterial species involved in cases of ovine respiratory complex that has been implicated to cause significant economic losses in sheep production system worldwide. The present study was undertaken with the aim of evaluating a SYBR Green dye based real time PCR assay targeting KMT1 gene for the detection of P. multocida. The analytical specificity and sensitivity of the PCR primers were evaluated. The test showed ten-fold more sensitivity than conventional PCR and detected down to 275.5 fg/ µl of genomic DNA concentration, equivalent to 100 copies of KMT1 gene of P. multocida. The real-time PCR was found to be specific for KMT1 gene of P. multocida, as no cross reactivity was detected with a variety of known bacterial isolates. A total of 52 ovine lung tissue samples were screened for P. multocida, which showed improved level of detection as compared to conventional PCR. It is concluded that, this assay may be used as a valuable diagnostic tool for the rapid and specific detection of P. multocida. By virtue of its high throughput format and its ability to accurately identify as well as quantify the bacterial DNA, the method may be useful in large scale epidemiological studies and clarification of pathogenesis.

Quantification of Pasteurella multocida in experimentally infected pigs using a real-time PCR assay

2017 ◽  
Vol 112 ◽  
pp. 177-184 ◽  
Author(s):  
V. Tocqueville ◽  
I. Kempf ◽  
F. Paboeuf ◽  
C. Marois-Créhan
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pasteurella Multocida ◽  
Pcr Assay

Feldvalidierung einer real-time PCR zum Nachweis von Mycoplasma hyopneumoniae in Nasentupfermaterial von lebenden Schweinen

2005 ◽  
Vol 147 (9) ◽  
pp. 373-379 ◽  
Author(s):  
F. Zeeh ◽  
P. Kuhnert ◽  
R. Miserez ◽  
M. G. Doherr ◽  
W. Zimmermann
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Mycoplasma Hyopneumoniae

Nutzen des hoch-sensitiven real-time-PCR HBV Tests (TaqMan) zur Prädiktion der Resistenzentwicklung bei Lamivudin-Langzeittherapie

2010 ◽  
Vol 48 (08) ◽  
Author(s):  
A Brodzinski ◽  
F van Bömmel ◽  
B Fülöp ◽  
B Schlosser ◽  
M Biermer ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr

Nachweis des porzinen Circovirus Typ 2 und des Torque-Teno-Sus-Virus 1 und 2 in Samenproben von Ebern einer österreichischen Besamungsstation

2011 ◽  
Vol 39 (04) ◽  
pp. 201-204
Author(s):  
A. Griessler ◽  
E. Pirker ◽  
H. Söllner ◽  
J. Segalés ◽  
T. Kekarainen ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Klinische Relevanz ◽  
Torque Teno Sus Virus

Zusammenfassung Gegenstand und Ziel: Das porzine Circovirus Typ 2 (PCV-2) und das Torque-teno-Sus-Virus (TTSuV) sind in schweineproduzierenden Ländern häufig nachzuweisen. Beide Erreger können sowohl horizontal als auch vertikal übertragen werden und Ebersamen könnte ein wichtiges Übertragungsmedium darstellen. Ziel der Studie war die Abklärung der Prävalenz dieser beiden Viren in Samenproben von Ebern. Material und Methoden: Von 100 Ebern einer Besamungsstation wurde jeweils eine Samenprobe mittels quantitativer Real-Time-PCR auf PCV-2 und mittels konventioneller PCR auf TTSuV-1 und TTSuV-2 untersucht. Ergebnisse: Nur bei einem Eber der Rasse Piétrain war ein positives PCV-2-Resultat festzustellen. TTSuV-1 ließ sich in vier Samenproben, TTSuV-2 in fünf Proben nachweisen. Ein Eber wies eine Koinfektion mit beiden TTSuV-Genotypen auf. Alle TTSuV-positiven Proben stammten von Piétrain-Ebern. Schlussfolgerung und klinische Relevanz: In der vorliegenden Studie wurde erstmals in Österreich TTSuV im Samen nachgewiesen. Die Prävalenz sowohl von TTSuV als auch von PCV-2 war gering. Die klinische Relevanz einer gleichzeitigen Kontamination des Samens mit beiden Viren ist nicht klar.

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