Novel PCR assay for the identification of two transmissible cancers in Cerastoderma edule

Transmissible cancers are a rare phenomenon in which cancer cells are passed between individuals, leading to the development of neoplasia in the host organism. Transmissible cancers have been identified in three independent clonal lineages in mammals and eight different clonal lineages in bivalves. This study focused on the development of a multiplex PCR assay for the detection of two types of bivalve transmissible neoplasias (BTN) of cockles Cerastoderma edule (CedBTN1 and CedBTN2). The diagnostic sensitivity (Se) and specificity (Sp) of the new PCR of hemolymph samples were assessed using a maximum likelihood estimation in the 88 samples compared to diagnoses obtained by histology and confirmed with genomic analyses. The results showed a Se of 100 % and a Sp of 98.1 % for CedBTN1, and a Se and Sp of 100 % for CedBTN2. The analytical Sp and Se were tested using DNA extractions from infected and non-infected C. edule and other bivalves affected by BTN. The assay demonstrated high analytical sensitivity and specificity, detecting down to 0.4 % of circulating CedBTN1 cells at a DNA concentration of 0.05 ng/µL in a hemolymph sample of a cockle with 85.7 % of CedBTN1 cells. The multiplex PCR assay was type-specific for CedBTN and capable of detecting both lineages simultaneously. Overall, this multiplex PCR assay is a reliable tool for the detection of transmissible cancers in cockles which will facilitate the diagnosis and monitoring of the disease.