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

Abstract

Lysozymes are used in sterilisation, antisepsis, dairy additives, inflammation, and cancer. One transgenic goat line expressing high levels of human lysozyme (hLZ) in goat milk has been developed in China. Herein, we established an event-specific real-time polymerase chain reaction (real-time PCR) method to detect the transgenic hLZ goat line. The developed method has high specificity, sensitivity and accuracy, and a wide quantitative dynamic range. The limit of detection and limit of quantification was 5 and 10 copies per reaction, respectively. The practical sample analysis results showed that the method could identify and quantify transgenic lysozyme content in trace samples in routine lab analyses. Furthermore, the potential applicability in risk assessment, such as molecular characterisation and gene horizontal transfer, was confirmed. We believe that this method is suitable for the detection of transgenic hLZ goat line and its derivate.

Development of an Event-Specific Droplet Digital PCR Assay for Quantification and Evaluation of the Transgene DNAs in Trace Samples of GM PRNP-Knockout Goat

“…Therefore, the LOQ was determined to be 7.2 copies HGE per reaction. The LOD and LOQ of ddPCR assay was lower than those of qPCR assays [ 7 , 12 , 13 ], indicating that the established KoP1 ddPCR assay is highly sensitive and has the potential to be applied in quantifying the KoP1 GM content in trace samples.…”

“…For example, one gene-specific qPCR method targeting Atlantic salmon ( Salmo salar L.) growth hormone gene ( GH1 ) was developed to quantify the S. salar L. DNA and Salmonidae ingredients in commercial foods [ 12 ]. We previously reported an event-specific qPCR method for quantifying the GM content of GM hLZ goat event [ 13 ]. However, there is still a need to construct the standard curves with series of calibrators for that qPCR analysis.…”