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Detection of Pasteurella multocida in experimentally infected embryonated chicken eggs by PCR assay
S B Shivachandra1, A A Kumar, R Gautam
1Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, India.
Abstract:
Applicability of polymerase chain reaction (PCR) assay to detect Pasteurella multocida in experimentally infected embryonated chicken egg was assessed in the present study. PCR assay rapidly and specifically detected the genome of P. multocida in amniotic fluid, allantoic fluid and homogenates of infected embryo and its membranes. The sensitivity of detection was as low as 20 bacterial cells/ml of allantoic or amniotic fluids. Detection of P. multocida in dead embryos by PCR was possible up to 6 and 30 days or more following storage of dead embryos at 37 degrees C, and at 4 degrees C as well as at -20 degrees C, respectively. The study revealed that PCR assays could be employed directly for detection and confirmation of P. multocida infection in experimentally infected chicken embryos.
Insights
Polymerase chain reaction (PCR) assay accurately detects Pasteurella multocida in chicken embryos. This rapid method identifies the bacteria in various embryo tissues, even after prolonged storage.
Area of Science:
- Veterinary Microbiology
- Molecular Diagnostics
- Avian Pathology
Background:
- Pasteurella multocida is a significant avian pathogen causing considerable economic losses in the poultry industry.
- Accurate and rapid diagnostic methods are crucial for controlling P. multocida outbreaks.
- Traditional diagnostic techniques can be time-consuming and lack sensitivity.
Purpose of the Study:
- To evaluate the applicability of the polymerase chain reaction (PCR) assay for detecting Pasteurella multocida in experimentally infected embryonated chicken eggs.
- To determine the sensitivity and specificity of the PCR assay.
- To assess the stability of P. multocida detection in stored infected embryos.
Main Methods:
- Experimental infection of embryonated chicken eggs with P. multocida.
- Sample collection from amniotic fluid, allantoic fluid, and embryo homogenates.
- Application of PCR assay for bacterial genome detection.
- Storage of infected embryos at different temperatures (37°C, 4°C, -20°C) for varying durations.
Main Results:
- The PCR assay demonstrated high specificity and rapid detection of P. multocida genome in all tested sample types.
- Sensitivity of the PCR assay was as low as 20 bacterial cells/ml.
- P. multocida was detectable in dead embryos for up to 6 days at 37°C and over 30 days at 4°C and -20°C.
Conclusions:
- The PCR assay is a reliable, sensitive, and specific tool for the direct detection and confirmation of P. multocida infection in experimentally infected chicken embryos.
- The method's ability to detect bacteria in stored samples enhances its practical utility in diagnostic laboratories.
- PCR offers a significant advancement over traditional methods for avian pasteurellosis diagnosis.
