Detection of cytomegalovirus in human placental cells by polymerase chain reaction.

Center for Medical Biology, Polish Academy of Sciences, Laboratory of Molecular Virology and Biological Chemistry, Lódź, Poland.
Apmis (Impact Factor: 1.92). 12/2006; 114(11):764-71. DOI: 10.1111/j.1600-0463.2006.apm_31.x
Source: PubMed

ABSTRACT Human cytomegalovirus (HCMV) is the most common cause of viral intrauterine infection. Progress in rapid, specific, and dependable detection of HCMV has recently been achieved by the use of DNA hybridization techniques and other molecular methods. We examined 21 placentas after delivery for the presence of HCMV DNA by polymerase chain reaction (PCR). To test the reliability of the PCR for the detection of HCMV DNA in clinical specimens, two simple PCR assays and a real-time quantitative PCR were used. PCR analysis of villous and decidual cells showed that HCMV DNA was present in 16 placentas (76.2%). Transmission of HCMV infection to chorionic villi was confirmed in 11 organs (52.4%), and congenital infections in newborns were detected in 9 cases (42.8%). These results suggest that HCMV genome detection in placentas at later gestational ages is common. Our results demonstrated that detection of HCMV DNA in placental tissues by DNA amplification provides a specific and sensitive method for diagnosis of intrauterine HCMV infection.

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    ABSTRACT: The association among specific single-nucleotide polymorphisms (SNPs) in TLR2 (Arg677Trp, Arg753Gln) and TLR4 (Asp299Gln) and human cytomegalovirus (CMV) infection was studied in infants and adults. The TLR2 and TLR4 polymorphisms were genotyped in 151 patients with CMV infections and in 78 unrelated healthy individuals. Genotyping was performed by restriction fragment length polymorphism (RFLP) analysis of PCR-amplified fragments. The viral load was measured by quantitative real-time PCR. Almost all of the patients with CMV infections were wild-type homozygotes without TLR2 and TLR4 polymorphisms. No significant differences in TLR2 and TLR4 polymorphisms were observed between infants with or without CMV infection. Compared with adults with CMV infections, heterozygosity for the TLR2 Arg677Trp and TLR4 Asp299Gly SNPs was detected more frequently in healthy individuals (p<0.05). Logistic regression analysis showed that the wild-type TLR2 genotype was associated with an increased risk of CMV infection and that heterozygosity for TLR2 and TLR4 SNPs diminished the risk of CMV infection in adult patients. An association between CMV load and the TLR4 SNP was found. Our results suggest that the wild-type TLR2 genotype may be a risk factor for CMV replication in adult patients.
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    ABSTRACT: Toll-like receptors (TLRs) play an important role in the subsequent initiation of innate and adaptive immune responses to microbial pathogens. Several single nucleotide polymorphisms (SNPs) within TLR genes seem to be associated with host-pathogen defense mechanisms. Little is known about the role of TLR polymorphisms in the pathogenesis of human cytomegalovirus (HCMV) infection. In the present study, we investigated the possible association between the polymorphisms of TLR2 (Arg677Trp and Arg753Gln), TLR4 (Asp299Gly) and TLR9 (T-1237C, T-1486C) with HCMV infection. Blood samples obtained from children and adults diagnosed with HCMV, and healthy control subjects were included in this study. The SNPs were genotyping by PCR-RFLP method. Digested PCR products were analysed by capillary electrophoresis. We did not observed differences in the frequencies of TLR2 (Arg753Gln), TLR4 and TLR9 genotypes among control and HCMV infected groups. Nonetheless, it is interesting that we found heterozygosity for the TLR2 Arg677Trp SNP in 72% health adults. In contrast, none of the HCMV-infected patients and children showed a heterozygous TLR2 polymorphism. In conclusion, our study demonstrated that SNPs in the TLR2 (Arg753Gln), TLR4 and TLR9 were not associated with HCMV infection.
    5th European Congress of Virology, Lyon, France; 09/2013


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Jun 30, 2014