Concurrent Hearing and Genetic Screening among Newborns in Ningbo, China.

OBJECTIVE: To detect the carrier rates of deafness gene variants in populations in Ningbo and analyze the risk of hereditary hearing loss through concurrent hearing and genetic screening tests. METHODS: Two thousand one hundred and seventy-four newborns were enrolled from November 2018 to August 2019. All subjects underwent hearing screening and newborn deafness genetic screening with 15 variants in 4 genes, and the positive sites were simultaneously verified by sequencing. RESULTS: The total carrier rate of genetic variants in Ningbo reached 4.32%, when GJB2 c.235delC was the variant with the highest prevalence (2.12%), approximately accounting for 48.9% of the total carrier frequency. The carrier frequency of SLC26A4 c.919-2A>G was 0.87%, while the most common variant in mitochondrial DNA (mtDNA) MT-RNR1 gene was m.1555A>G, and its carrier frequency was 0.184%. In the OAE testing, 92 newborns passing hearing screening were tested positively for variants in 4 genes, and 2 of 42 newborns who failed in the first hearing test were found to mutate in 4 genes. CONCLUSION: Herein, the results concerning the carrier rates for deafness gene mutations of Ningbo population are reported. Our study is beneficial to the insight into the deafness genomic epidemiology for deafness genes in Ningbo population and provides the reference for healthcare in Ningbo.

Targeted identification of C-type lectins in snake venom by 2DE and Western blot.

C-type lectins (CTL) and CTL-like proteins (snaclecs) are important toxins found in snake venom which can disrupt hemostasis by binding platelet membrane glycoproteins. Traditional identification of these toxins usually relies on an "activity-directed fractionation" approach which is very arduous. Here, we report a new method for rapid screening of these proteins in snake venom. METHODS: A conserved and immunogenic peptide found in svCTLs (CTL and snaclecs) was identified by sequence alignment using DNAStar software. The peptide was de novo synthesized and conjugated to keyhole limpet hemocyanin (KLH). Rabbit antibodies were generated against the peptide by classical immunization. Deinagkistrodon acutus venom was separated by two-dimensional electrophoresis (2DE) followed by Western blot and CTLs immunodetected using the isolated polyclonal antibody. The same svCTL spots on a parallel 2DE gel were isolated and analyzed by MALDI-TOF-MS. RESULTS: A highly conserved peptide with the sequence "KTWDDAEKFCTEQ" was identified as a common epitope in svCTLs. The polyclonal antibody against the 13aa-peptide was successfully prepared and purified. Its usefulness to detect svCTLs in D. acutus venom was tested by 2DE-WB and we determined that it positively identified all known D. acutus venom CTLs. CONCLUSIONS: Immunodetection with antibodies against KTWDDAEKFCTEQ is an efficient strategy to identify novel svCTLs in the context of a complex proteome.