MIPs: multi-locus intron polymorphisms in species identification and population genomics.

The study of species groups in which the presence of interspecific hybridization or introgression phenomena is known or suspected involves analysing shared bi-parentally inherited molecular markers. Current methods are based on different categories of markers among which the classical microsatellites or the more recent genome wide approaches for the analyses of thousands of SNPs or hundreds of microhaplotypes through high throughput sequencing. Our approach utilizes intron-targeted amplicon sequencing to characterise multi-locus intron polymorphisms (MIPs) and assess genetic diversity. These highly variable intron regions, combined with inter-specific transferable loci, serve as powerful multiple-SNP markers potentially suitable for various applications, from species and hybrid identification to population comparisons, without prior species knowledge. We developed the first panel of MIPs highly transferable across fish genomes, effectively distinguishing between species, even those closely related, and populations with different structures. MIPs offer versatile, hypervariable nuclear markers and promise to be especially useful when multiple nuclear loci must be genotyped across different species, such as for the monitoring of interspecific hybridization. Moreover, the relatively long sequences obtained ease the development of single-locus PCR-based diagnostic markers. This method, here demonstrated in teleost fishes, can be readily applied to other taxa, unlocking a new source of genetic variation.

Evaluation of the immune response to COVID-19 vaccine mRNA BNT162b2 and correlation with previous COVID-19 infection.

BACKGROUND: The kinetics of immune response after vaccination with mRNA-BNT162b2 (Comirnaty®) and the correlation with previous COVID-19 infection are still unclear. METHODS: Thirty-six subjects receiving mRNA-BNT162b2 were prospectively studied [10 days after the first dose (Time 1), 7 days and 16 weeks after the second dose (Time 2 and Time 3)] to determine antibody titers against nucleocapside, trimeric spike protein (TSP) and receptor-binding-domain (RBD) of the spike protein. Ten subjects had a previous COVID-19 infection not requiring hospitalization (Group 1) and 26 did not (Group 2). RESULTS: At Time 1 all subjects in Group 1 had IgG against TSP > 800 AU/mL compared to 11/26 (42.3%) in Group 2, whilst at Time 2 all subjects in both groups had > 800 AU/mL. The mean IgG against TSP titer at Time 3 was 711 AU/mL (95% CI 652-800) in Group 1 and 240 AU/mL (95% CI 112-375) in Group 2 (p < 0.0001). However, all subjects in both groups maintained antibody titers above the lower threshold limit at each time-point considered. These results were confirmed also using anti-RBD antibodiy tests. Antibodies against nucleocapside were reactive only in subjects in Group 1 and remained stable during the study period. No subject had a new onset of COVID-19 infection within 16 weeks of follow-up. CONCLUSIONS: Subjects with previous COVID-19 infection have a more rapid immune response to mRNA-BNT162b2 than others and maintained higher antibody titers during 16 weeks of follow-up. However, no new COVID-19 infection also in subjects with lower antibody titers.