The invention discloses a multichannel fluorescent PCR detection kit for a congenital deafness gene. With 35delG, 155delTCTG,176dell6bp, 235delC, 299-300delAT and 512insAACG mutations of a GJB2 gene, and 538G>T and 547G>A mutations of a GJB3 gene as detection objects, specific primers and a probe are designed, wherein the probe is used for specifically detecting whether 35, 155, 176, 235 and 299 loci of the GJB2 gene, and 538 and 547 loci of the GJB3 gene contain mutations or not. According to the multichannel fluorescent PCR detection kit, eight mutation loci of the congenital deafness are detected at the same timefor the first time in a same reaction tube by applying a multichannel fluorescent PCR method; the operation is simple, convenient, fast, accurate, high in flux, and low in cost; cross contamination is avoided by a pipe closing operation; and the deafness screening is high in intentionality and wide in screening range.