P2/P8 primer PCR effective in sex determination in Domestic birds

Abstract

In avian species, sex determination is based on ZW-ZZ chromosomes. Males are homogametic have two Z chromosomes (ZZ), while females are heterogametic and have a Z and a W chromosome (ZW). The PCR has been successfully used for the detection of the sex in birds. In the current study, an effort to compare the accuracy of sex determination using primers P2/P8 & P2/NP/MP, targeting the Chromo-Helicase DNA binding (CHD) gene, in Domestic birds of known gender using PCR technique. A total of seventy-four sexed samples were identified by the P2/P8 PCR. A total of 72 valid results were obtained while two samples failed to provide an amplification. The female (ZW) showed two bands (350 & 365 bp approximately) separated from each other by a mere 15 bp, while, males (ZZ) showed a single band at 350 bp. Out of the seventy-two results all seventy-two samples were correctly identified by the test. The overall accuracy of the test stood at 97.29% (72/74).In the P2/NP/MP PCR, the female (ZW) showed two bands (330 & 270 bp) separated from each other by a 60 bp, while, males (ZZ) showed a single band at 270 bp. A total of 74 valid results were obtained. Out of the 74 samples sexed using P2/NP/MP PCR test, all 74 samples provided an amplification. However, out of the seventy-four samples, seven samples provided erroneous results. The overall accuracy of the test stood at 90.54%.Thus, the P2/P8 primer was more accurate in the sex estimation in domestic birds, whereas, the P2/MP/NP PCR has comparatively less accurate in detecting the sex in domestic birds. Although, it is worth mentioning that the P2/MP/NP PCR was 100% accurate in the estimation of sex in Khaki campbell duck (Anas platyrhynchos domesticus), Guinea fowl (Numida meleagris), Turkey (Meleagris gallopavo) and Japanese quail (Coturnix japonica). Depending on targeted species the P2/P8 PCR can be used to identify birds with precision.