An Event-Specific Real-Time PCR Method for Measuring Transgenic Lysozyme Goat Content in Trace Samples

Area of Science:

• Biotechnology
• Molecular Biology
• Genetics

Background:

• Lysozymes have diverse applications including sterilization, antisepsis, and in the dairy industry.
• A transgenic goat line expressing high levels of human lysozyme (hLZ) has been developed.
• Accurate detection methods are needed for monitoring transgenic organisms.

Purpose of the Study:

• To establish an event-specific real-time polymerase chain reaction (real-time PCR) method.
• To detect and quantify the transgenic hLZ goat line.
• To assess the method's applicability in routine analyses and risk assessment.

Main Methods:

• Development of an event-specific real-time PCR assay.
• Validation of specificity, sensitivity, accuracy, and dynamic range.
• Testing with practical sample analysis.

Main Results:

• The real-time PCR method demonstrated high specificity, sensitivity, and accuracy.
• The limit of detection was 5 copies/reaction, and the limit of quantification was 10 copies/reaction.
• The method successfully identified and quantified hLZ in trace samples.

Conclusions:

• The developed real-time PCR method is suitable for detecting the transgenic hLZ goat line.
• The method can be applied in routine laboratory analyses.
• Potential applicability in risk assessment, including molecular characterization and gene horizontal transfer, was confirmed.