Ocena związku poziomu wiremii HCMV u matki z przebiegiem ciąży i stanem urodzeniowym noworodków

Congenital cytomegaly is caused by intrauterine mother-to-fetus HCMV transmission and constitutes the most common vertical infection. Objectives: The aim of the study was to analyze the viremia level in maternal blood and its influence on the course and duration of pregnancy, as well as newborn condition. Material and methods: The material included blood samples collected from 117 pregnant women with serological features of HCMV infection and from 29 neonates hospitalized at DFMMG in Lodz between 1999 and 2009. The presence of HCMV DNA in the maternal and fetal blood was tested using real-time PCR. Results: Prevalence of maternal viremia was observed to increase the risk of viremia in neonates, as compared to children born to mothers with no viremia. However, lack of HCMV DNA in maternal blood does not exclude fetal infection in utero. Newborn condition assessed by Apgar scores was significantly lower in the group of infants born to mothers with serological features of acute cytomegaly (pWrodzona cytomegalia wywołana transmisją wirusa cytomegalii (HCMV) od matki do płodu, jest najczęstszym zakażeniem wertykalnym. Cel pracy: Celem pracy była ocena poziomu wiremii u kobiet ciężarnych i jej wpływu na przebieg i czas trwania ciąży oraz stan urodzeniowy noworodków. Materiał i metody: Materiałem do badań była krew pobrana od 117 ciężarnych z serologicznymi cechami zakażenia HCMV oraz 29 noworodków hospitalizowanych w Klinice Medycyny Matczyno-Płodowej i Ginekologii Instytutu Centrum Zdrowia Matki Polki w latach 1999-2009. Liczba kopii DNA HCMV we krwi matek i dzieci oznaczana była metodą PCR w czasie rzeczywistym (real-time PCR). Wyniki: Stwierdzono, że występowanie wiremii HCMV u matki zwiększa ryzyko występowania wiremii u noworodków w porównaniu z ryzykiem u dzieci matek bez wiremii, jednakże brak DNA HCMV we krwi matki nie wyklucza zakażenia płodu in utero. Stan urodzeniowy noworodków oceniany w skali Apgar był istotnie niższy w grupie noworodków urodzonych przez matki z serologicznymi cechami ostrej cytomegalii (

TLR9 -1486T/C and 2848C/T SNPs Are Associated with Human Cytomegalovirus Infection in Infants.

Toll-like receptor 9 (TLR9) recognizes non-methylated viral CpG-containing DNA and serves as a pattern recognition receptor that signals the presence of human cytomegalovirus (HCMV). Here, we present the genotype distribution of single-nucleotide polymorphisms (SNPs) of the TLR9 gene in infants and the relationship between TLR9 polymorphisms and HCMV infection. Four polymorphisms (-1237T/C, rs5743836; -1486T/C, rs187084; 1174G/A, rs352139; and 2848C/T, rs352140) in the TLR9 gene were genotyped in 72 infants with symptomatic HCMV infection and 70 healthy individuals. SNP genotyping was performed by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Digested fragments were separated and identified by capillary electrophoresis. The HCMV DNA copy number was measured by a quantitative real-time PCR assay. We found an increased frequency of heterozygous genotypes TLR9 -1486T/C and 2848C/T in infants with HCMV infection compared with uninfected cases. Heterozygous variants of these two SNPs increased the risk of HCMV disease in children (P = 0.044 and P = 0.029, respectively). In infants with a mutation present in at least one allele of -1486T/C and 2848C/T SNPs, a trend towards increased risk of cytomegaly was confirmed after Bonferroni's correction for multiple testing (Pc = 0.063). The rs352139 GG genotype showed a significantly reduced relative risk for HCMV infection (Pc = 0.006). In contrast, the -1237T/C SNP was not related to viral infection. We found no evidence for linkage disequilibrium with the four examined TLR9 SNPs. The findings suggest that the TLR9 -1486T/C and 2848C/T polymorphisms could be a genetic risk factor for the development of HCMV disease

Demographic and clinical characteristics of study subjects with HCMV infection.

<p>Demographic and clinical characteristics of study subjects with HCMV infection.</p

The distribution of allele frequencies of <i>TLR9</i> SNPs in infants with and without HCMV infection.

<p>The distribution of allele frequencies of <i>TLR9</i> SNPs in infants with and without HCMV infection.</p

Comparison of viremia levels in HCMV-infected infants without or with the <i>TLR9</i> -1486T/C SNP (N = 72).

<p>Bars in the scatter dot plot represent the mean viral loads. Bars in the box plots represent median viral loads, upper and lower borders represent the 25th and 75th percentiles, and whiskers represent the minimum to maximum values. <i>P</i> ≤ 0.05, Mann-Whitney U test.</p

Visualization of selected PCR-RFLP products for <i>TLR9</i> -1486T/C genotyping.

<p>Gel image: 1 and 2, heterozygous TC genotype (192, 327, 490 bp); 3 and 4, TT genotype (192, 327 bp); 5 and 6, CC genotype (490 bp). Alignment markers (15 bp, 1 kbp).</p

<i>TLR9</i> SNP variance from Hardy-Weinberg equilibrium.

<p><i>TLR9</i> SNP variance from Hardy-Weinberg equilibrium.</p

Restriction enzymes and length of the restriction fragments.

<p>Restriction enzymes and length of the restriction fragments.</p

The distribution of genotypes frequencies of <i>TLR9</i> SNPs in infants and relationship between polymorphisms and the risk of HCMV infection.

<p>The distribution of genotypes frequencies of <i>TLR9</i> SNPs in infants and relationship between polymorphisms and the risk of HCMV infection.</p