Impact of human milk banking on neonatal mortality, necrotizing enterocolitis, and exclusive breastfeeding - experience from a tertiary care teaching hospital, south India.

OBJECTIVE: The objective of this study is to study the impact of a Human Milk Bank (HMB) on neonatal mortality, incidence of necrotizing enterocolitis (NEC) and rate of exclusive breastfeeding. METHODS: This pre-post intervention study was conducted in a tertiary care teaching institute in south India. Data regarding neonatal mortality, incidence of NEC, and exclusive breastfeeding rates were collected for a period of 6 months before and after establishing a modern HMB and compared. RESULTS: The number of deliveries, live births, and incidence of preterm and VLBW neonates during pre- and post-HMB periods were comparable. Neonatal mortality was 11.32/1000 live births pre-HMB compared with 10.77/1000 live births post HMB. The incidence of NEC was 1.26% of live births pre-HMB compared with 1.07% post-HMB. Exclusive breastfeeding rate pre-HMB was 34% compared with 74% post HMB (p < .001). CONCLUSION: There is a decreasing trend in neonatal mortality and incidence of NEC after establishing a HMB. Human milk banking significantly improved exclusive breastfeeding rate in the population studied.

Modified low cost SNP genotyping technique using cycle threshold (Ct) & melting temperature (Tm) values in allele specific real-time PCR.

BACKGROUND & OBJECTIVES: Genotyping has now become one of the major diagnostic means for almost all diseases. Among the advanced techniques that are used to study single nucleotide polymorphisms (SNPs), only a few are applicable for routine disease diagnosis. Their applicability mainly depends on three factors: cost, time, and accuracy. The primary objective of this study was to propose allele-specific real-time PCR as a rapid, low cost and simple genotyping method for routine diagnostics. METHODS: Two SNPs, rs3014866 and rs2149356 were analysed using allele-specific real-time PCR. The polymerase chain reaction was carried out using RealQ PCR master mix containing SYBR Green DNA I dye followed by melt curve analysis. The results were validated by agarose gel electrophoresis and DNA sequencing. RESULTS: The allelic discrimination and zygosity of the two SNPs were assessed by combined cycle threshold (Ct) and melting temperature (T m ) values. Variations in Ct and T m values among the two alleles were observed in both rs3014866 (Ct: C allele - 24 ± 1, T allele - 27 ± 1; T m: C allele - 82.5 ± 0.3, T allele - 86.3 ± 0.2) and rs2149356 (Ct: C allele - 24 ± 1, A allele - 26 ± 1; T m: C allele - 79.4 ± 0.2, A allele - 80.4 ± 0.3). Based on the variations, homozygous and heterozygous alleles were detect ed. Agarose gel electrophoresis and DNA sequencing also confirmed the allelic variation and zygosity observed in real-time PCR. INTERPRETATION & CONCLUSIONS: In diagnostic settings where a large number of samples are analysed daily, allele-specific real-time PCR assay may serve as a simple, low cost and efficient method of genotyping.