Genetic variation in the MBL2 gene is associated with Chlamydia trachomatis infection and host humoral response to Chlamydia trachomatis infection

This study aims to assess the potential association of MBL2 gene single nucleotide polymorphisms (SNPs) to Chlamydia trachomatis infection. We analysed a selected sample of 492 DNA and serum specimens from Dutch Caucasian women. Women were categorized into four groups of infection status based on the results of DNA and antibody tests for C. trachomatis: Ct-DNA+/IgG+, CtDNA+/IgG−, Ct-DNA−/IgG+, and Ct-DNA−/IgG−. We compared six MBL2 SNPs (−619G > C (H/L), −290G > C (Y/X), −66C > T (P/Q), +154C > T (A/D), +161A > G (A/B), and +170A > G (A/C)) and their respective haplotypes in relation to these different subgroups. The −619C (L) allele was less present within the Ct-DNA−/IgG+ group compared with the Ct-DNA−/IgG− group (OR = 0.49; 95% CI: 0.28–0.83), while the +170G (C) allele was observed more in the Ct-DNA+/IgG+ group as compared with the Ct-DNA−/IgG− group (OR = 2.4; 95% CI: 1.1–5.4). The HYA/HYA haplotype was more often present in the Ct-DNA−/IgG− group compared with the Ct-DNA+/IgG+ group (OR = 0.37; 95% CI: 0.16–0.87). The +170G (C) allele was associated with increased IgG production (p = 0.048) in C. trachomatis PCR-positive women. This study shows associations for MBL in immune reactions to C. trachomatis. We showed clear associations between MBL2 genotypes, haplotypes, and individuals’ stages of C. trachomatis DNA and IgG positivity.NGI Life Sciences Pre-Seed and a EuroTransBio grant.https://www.mdpi.com/journal/ijerphMedical Microbiolog

Genetic Variation in the Gene Is Associated with Infection and Host Humoral Response to Infection.

This study aims to assess the potential association of MBL2 gene single nucleotide polymorphisms (SNPs) to Chlamydia trachomatis infection. We analysed a selected sample of 492 DNA and serum specimens from Dutch Caucasian women. Women were categorized into four groups of infection status based on the results of DNA and antibody tests for C. trachomatis: Ct-DNA+/IgG+, Ct-DNA+/IgG-, Ct-DNA-/IgG+, and Ct-DNA-/IgG-. We compared six MBL2 SNPs (-619G > C (H/L), -290G > C (Y/X), -66C > T (P/Q), +154C > T (A/D), +161A > G (A/B), and +170A > G (A/C)) and their respective haplotypes in relation to these different subgroups. The -619C (L) allele was less present within the Ct-DNA-/IgG+ group compared with the Ct-DNA-/IgG- group (OR = 0.49; 95% CI: 0.28-0.83), while the +170G (C) allele was observed more in the Ct-DNA+/IgG+ group as compared with the Ct-DNA-/IgG- group (OR = 2.4; 95% CI: 1.1-5.4). The HYA/HYA haplotype was more often present in the Ct-DNA-/IgG- group compared with the Ct-DNA+/IgG+ group (OR = 0.37; 95% CI: 0.16-0.87). The +170G (C) allele was associated with increased IgG production (p = 0.048) in C. trachomatis PCR-positive women. This study shows associations for MBL in immune reactions to C. trachomatis. We showed clear associations between MBL2 genotypes, haplotypes, and individuals' stages of C. trachomatis DNA and IgG positivity

Genetic Variation in the Gene Is Associated with Infection and Host Humoral Response to Infection.

This study aims to assess the potential association of MBL2 gene single nucleotide polymorphisms (SNPs) to Chlamydia trachomatis infection. We analysed a selected sample of 492 DNA and serum specimens from Dutch Caucasian women. Women were categorized into four groups of infection status based on the results of DNA and antibody tests for C. trachomatis: Ct-DNA+/IgG+, Ct-DNA+/IgG-, Ct-DNA-/IgG+, and Ct-DNA-/IgG-. We compared six MBL2 SNPs (-619G > C (H/L), -290G > C (Y/X), -66C > T (P/Q), +154C > T (A/D), +161A > G (A/B), and +170A > G (A/C)) and their respective haplotypes in relation to these different subgroups. The -619C (L) allele was less present within the Ct-DNA-/IgG+ group compared with the Ct-DNA-/IgG- group (OR = 0.49; 95% CI: 0.28-0.83), while the +170G (C) allele was observed more in the Ct-DNA+/IgG+ group as compared with the Ct-DNA-/IgG- group (OR = 2.4; 95% CI: 1.1-5.4). The HYA/HYA haplotype was more often present in the Ct-DNA-/IgG- group compared with the Ct-DNA+/IgG+ group (OR = 0.37; 95% CI: 0.16-0.87). The +170G (C) allele was associated with increased IgG production (p = 0.048) in C. trachomatis PCR-positive women. This study shows associations for MBL in immune reactions to C. trachomatis. We showed clear associations between MBL2 genotypes, haplotypes, and individuals' stages of C. trachomatis DNA and IgG positivity

Analysis of multiple single nucleotide polymorphisms (SNP) on DNA traces from plasma and dried blood samples

Reliable analysis of single nucleotide polymorphisms (SNPs) in DNA derived from samples containing low numbers of cells or from suboptimal sources can be difficult. A new procedure to characterize multiple SNPs in traces of DNA from plasma and old dried blood samples was developed. Six SNPs in the Mannose Binding Lectin 2 (MBL2) gene were chosen as targets for analysis. DNA was extracted from plasma obtained from mothers (n=49) and their neonates (n=49) and from old dried blood samples (n=204). Multiple Real-Time SNP analyses in the MBL2 gene were carried out on all samples. Because of very low DNA concentrations in most of the samples, a pre-amplification step was utilized. It was possible to analyze all plasma samples (n=98), including those with very low cell numbers (n=21) and 93% of the old dried blood samples (n=189). Results obtained from pre-amplified samples were in full agreement with neat samples. All possible SNP alleles were present in our population. The frequencies of the different alleles from both plasma and dried blood samples (n=287) were in agreement with earlier studies of the Caucasian population. In conclusion, amplification prior to Real-Time PCR SNP analysis is a convenient, cost effective and useful method to significantly improve the reliable SNP detection in specimens containing very low concentrations or poor quality DNA from suboptimal sources

Mannose-binding lectin in term newborns and their mothers: genotypic and phenotypic relationship

Functional mannose-binding lectin (f-MBL) plays an important role in the innate neonatal immune system. We studied the origin of f-MBL in umbilical cord blood (UCB) by measuring maternal MBL (n=47), collected before elective cesarean section, and neonatal MBL (n=43) in arterial umbilical cord blood. In a subgroup, arterial and venous UCB MBL levels were measured. In addition, MBL expression was correlated with genetic mutations. The f-MBL levels in term infants were lower than in their mothers (0.70 microg/ml vs 1.11 microg/ml, p <0.01) and maternal and neonatal MBL levels were only weakly correlated (R=0.32, p <0.001), which suggests a fetal origin of f-MBL. Arterial and venous UCB median MBL levels did not differ (0.98 microg/ml vs. 1.40 microg/ml, p=0.20). No homozygous mutations were found. MBL was lower in mothers and infants with a (compound) heterozygous mutation than in those with a wild type. One new (HYPB) and two rare haplotypes (HXPA, LYPD) were reported in our population. Levels of MBL differed depending on the genotype of the mother or the infant. Because the role of MBL in host defense is still unclear, both f-MBL and haplotype should be measured to determine the clinical implications of MBL deficiency in infant

Flowchart of experimental set-up.

<p>SA, <i>S. aureus</i>; PA, <i>P. aeruginosa</i>; CA, <i>C. albicans</i>; CFU, colony forming unit.</p

Overview of primers and probes used for pathogen detection.

<p>Overview of primers and probes used for pathogen detection.</p

Detection rates (percentage of positive PCRs) of 3 different DNA isolation methods in dilutions series of 1–1000 CFU/ml.

<p>Fisher’s exact test performed on 1 CFU/ml samples, statistically significant when <i>p</i>≤0.05. a; Polaris versus MolYsis, b; Polaris versus TTE-EasyMAG, c; MolYsis versus TTE-EasyMAG.</p

Comparison of Polaris and MolYsis methods using 1 ml and 5 ml spiked whole blood samples.

<p>The grey bars represent the Polaris samples (1 or 5 ml whole blood), and the white bars represent the MolYsis isolated samples (1 or 5 ml whole blood). SEM is shown. The numbers in the bars represent the sample numbers.</p