Novel LNA probe-based assay for the A1 and A2 identification of beta-casein gene in milk samples

Author(s)

Giglioti, Rodrigo ; Okino, Cintia Hiromi ; Azevedo, Bianca Taina ; Gutmanis, Gunta ; Katiki, Luciana Morita ; de Sena Oliveira, Marcia Cristina ; Vercesi Filho, Anibal Eugenio

• Document type: Journal article
• Source: FOOD CHEMISTRY: MOLECULAR SCIENCES; v. 3, p. 6-pg., 2021-12-30.

Abstract

The rising consumption of A2 milk and its derivatives in recent years has garnered attention from both consumers and producers, mainly due its possible health benefits, such as enhanced digestion and easier absorption. Thus, a novel real-time PCR using a combination of locked nucleic acid modified (LNA) conjugated probes was developed to genotype A1 and A2 alleles of beta-casein gene (CSN2) and to detect and quantify the A1 presence in A2 samples. The limit of detection for each probe (A1 and A2) was evaluated using decreasing serial dilutions. Besides, the sensitivity of A1 allele detection in the A2 samples was also tested. The limits of detection of A1 and A2 alleles were 6 copies, while for A1 allele detection in A2 samples was 7.5 copies (1%). The LNA-probe based method was found to be rapid, robust, highly sensitive, cost effective, and can be employed as screening test to certificate the A2 dairy products. (AU)

• FAPESP's process: 18/19452-2 - Multi user equipment approved in grant 2017/50.339-5: nanoespectrofotômetro BioDrop
• Grantee: Renata Helena Branco Arnandes
• Support Opportunities: Multi-user Equipment Program