Wujcicka Wioletta, Paradowska Edyta, Studzińska Mirosława, Gaj Zuzanna, Wilczyński Jan, Leśnikowski Zbigniew, Nowakowska Dorota
Scientific Laboratory of Center of Medical Laboratory Diagnostics, Polish Mother's Memorial Hospital-Research Institute, Lodz, Poland.
Laboratory of Molecular Virology and Biological Chemistry, Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland.
PLoS One. 2015 Apr 6;10(4):e0122831. doi: 10.1371/journal.pone.0122831. eCollection 2015.
Some single nucleotide polymorphisms (SNP), located in Toll-like receptor (TLR) genes, were reported to be associated with human cytomegalovirus (HCMV) infections. The study was aimed to assess the correlation of SNPs at TLR4 and TLR9 genes with the occurrence of congenital cytomegaly, based on available samples.
Reported case-control study included both HCMV infected and non-infected fetuses and newborns. The specimens were classified to the molecular analyses, based on serological features of the recent infection and HCMV DNAemia in body fluids. TLR SNPs were studied, using multiplex nested PCR-RFLP assay, and determined genotypes were confirmed by sequencing. Hardy-Weinberg equilibrium was assessed for the identified genotypes. The linkage disequilibrium was also estimated for TLR4 SNPs. A relationship between the status of TLR genotypes and congenital cytomegaly development was estimated, using a logistic regression model.
Hardy Weinberg equilibrium was observed for almost all SNPs, both infected and non-infected patients, with exception of TLR4 896 A>G polymorphism in the control group (P≤0.050). TLR4 896 A>G and 1196 C>T SNPs were found in linkage disequilibrium in both study groups (P≤0.050). The CC genotype at TLR4 1196 SNP and the GA variant at TLR9 2848 G>A SNP were significantly associated with HCMV infection (P≤0.050). The risk of congenital cytomegaly was higher in heterozygotes at TLR9 SNP than in the carriers of other genotypic variants at the reported locus (OR 4.81; P≤0.050). The GC haplotype at TLR4 SNPs and GCA variants at TLR4 and TLR9 SNPs were significantly associated with HCMV infection (P≤0.0001). The ACA variants were more frequent among fetuses and neonates with symptomatic, rather than asymptomatic cytomegaly (P≤0.0001).
TLR4 and TLR9 polymorphisms may contribute to the development of congenital infection with HCMV in fetuses and neonates. The TLR9 2848 GA heterozygotic status possibly predisposes to HCMV infection, increasing the risk of congenital cytomegaly development.
据报道，位于Toll样受体（TLR）基因中的一些单核苷酸多态性（SNP）与人类巨细胞病毒（HCMV）感染相关。本研究旨在基于现有样本评估TLR4和TLR9基因中的SNP与先天性巨细胞病毒感染发生情况的相关性。
所报道的病例对照研究纳入了HCMV感染和未感染的胎儿及新生儿。根据近期感染的血清学特征和体液中的HCMV血症，将标本分类进行分子分析。采用多重巢式PCR-RFLP分析法研究TLR SNP，并通过测序确认所确定的基因型。对所鉴定的基因型评估哈迪-温伯格平衡。还对TLR4 SNP估计连锁不平衡。使用逻辑回归模型评估TLR基因型状态与先天性巨细胞病毒感染发展之间的关系。
几乎所有SNP在感染和未感染患者中均观察到哈迪-温伯格平衡，但对照组中的TLR4 896 A>G多态性除外（P≤0.050）。在两个研究组中均发现TLR4 896 A>G和1196 C>T SNP处于连锁不平衡状态（P≤0.050）。TLR家族4 1196 SNP的CC基因型和TLR9 2848 G>A SNP的GA变异与HCMV感染显著相关（P≤0.050）。TLR9 SNP杂合子发生先天性巨细胞病毒感染的风险高于所报道位点其他基因型变异的携带者（比值比4.81；P≤0.050）。TLR4 SNP的GC单倍型以及TLR4和TLR9 SNP的GCA变异与HCMV感染显著相关（P≤0.0００1）。ACA变异在有症状而非无症状巨细胞病毒感染的胎儿和新生儿中更为常见（P≤0.0００1）。
TLR4和TLR9多态性可能有助于胎儿和新生儿先天性HCMV感染的发展。TLR9 2848 GA杂合状态可能易患HCMV感染，增加先天性巨细胞病毒感染发展的风险。
