Cytomegalovirus genotype distribution among postnatally infected infants: association of glycoprotein B, glycoprotein N and glycoprotein H types with CMV-associated thrombocytopenia

The aim of the present study was to identify the virulence of glycoprotein B (gB), glycoprotein N (gN), and glycoprotein H (gH) genotypes of cytomegalovirus(CMV) and assess possible relationships between genotypes and CMV-associated thrombocytopenia. CMV gB, gN, and gH strains were determined by nested PCR and restriction length polymorphism from 30 CMV-associated thrombocytopenia infants infected postnatally and 40 non-thrombocytopenic infants. The gN2 (p = 0.043) and gH2 (p = 0.038) genotypes were associated with an elevated risk of developing thrombocytopenia. gB1 genotype was detected in 80.0% (16/20) of infants with moderately to severely symptomatic CMV disease and was associated with severe manifestations in CMV-associated thrombocytopenia infants (p = 0.022). Conversely, the gN1 genotype was detected in 5.0% (1/20) of infants with moderately to severely symptomatic CMV disease and represent less pathogenic CMV strains (p = 0.044). There may be potential associations between the gN2 and gH2 genotypes of CMV and infantile thrombocytopenia, and that the detection of the gB1and gN1 genotypes may help define the severity of CMV disease in infants.

Genetic characterization ofToxoplasma gondiiisolates from Portugal, Austria and Israel reveals higher genetic variability within the type II lineage

SUMMARYThis study compared genetic diversity ofToxoplasma gondiiisolates from Portugal, Austria and Israel. For this, we genotyped 90T. gondiiisolates (16 from Portugal, 67 from Austria and 7 from Israel) using 10 nested PCR-restriction length polymorphism (RFLP) genetic markers and 15 microsatellite (MS) markers. By PCR-RFLP typing, 7 isolates from Portugal chickens were identified as type II (ToxoDB #1 or #3), 4 were type III (ToxoDB #2) and the remaining 4 isolates have unique genotype pattern were designated as ToxoDB #254. One mouse virulent isolate from a bovine fetus (Bos taurus) in Portugal was type I (ToxoDB #10) at all loci and designated as TgCowPr1. All 67 isolates from Austria and 7 from Israel were type II (ToxoDB #1 or #3). By MS typing, many additional genetic variations were revealed among the type II and type III isolates. Phylogenetic analysis showed that isolates from the same geographical locations tend to cluster together, and there is little overlapping of genotypes among different locations. This study demonstrated that the MS markers can provide higher discriminatory power to reveal association of genotypes with geographical locations. Future studies of the type II strains in Europe by these MS markers will be useful to reveal transmission patterns of the parasite.

Distribution of Angiotensin-1 Converting Enzyme Insertion/Deletion and α-Actinin-3 Codon 577 Polymorphisms in Turkish Male Soccer Players

Angiotensin-1 converting enzyme (ACE) gene and α-actinin-3 (ACTN3) gene polymorphisms are considered to be the most important candidate genes for genetic predisposition to human athletic performance. In the present study, we aimed to analyze the distribution of ACE and ACTN3 polymorphisms for the first time in male Turkish soccer players. In this prospective study, our cohort consisted of 25 professional players, all with Turkish ancestry. Polymerase chain reaction (PCR)-restriction length polymorphism was used for the characterization of the genotype of ACTN3 and single PCR for ACE. For ACE genotype, 16%, 44%, and 40% of the players had insertion/insertion (II), insertion/deletion (ID), and deletion/deletion (DD) genotypes, respectively, whereas 20% had XX, 36% had RX, and 44% had RR genotypes for ACTN3. When we examined the allelic percentages, for ACE, D allele was recorded as 62 and I as 38, and for ACTN3, R allele was 62 and X was 38. Our results were in agreement with the previous reports, indicating the presence of ACTN3 D and ACE X allele in soccer players. We suggest that ACE and ACTN3 genotypes are important biomarkers for genetic counseling for the individuals who are prone to be successful soccer players.

Molecular methods for the detection and characterization of foodborne pathogens

The surveillance of foodborne pathogens in food industries has shown the urgent need for rapid and dependable methods to detect and characterize the organisms in food and environments of clinical and epidemiologic importance. Recent studies on rapid methods in microbiology have been focused on biochemical characterization, immunoassays, and molecular methods. Many molecular methods have been developed and applied to the detection and characterization of foodborne pathogens in laboratories and food industries. They can be mainly divided into DNA banding pattern-based tests and DNA sequence-based tests. The former includes nucleic acid hybridization, polymerase chain reaction (PCR), amplified restriction length polymorphism, and randomly amplified polymorphic DNA, etc. Most of these methods in commercial applications are based on PCR or hybridization techniques. The principle, characteristics, and application of molecular methods for the detection and characterization of foodborne pathogens were reviewed in this article.

PTPN22 Mutation Is Present in An Increased Proportion of ITP Patients.

Immune-mediated thrombocytopenic purpura (ITP) is an autoimmune disorder characterized by low platelet count and antibody mediated platelet destruction. ITP may be due to a failure of T-cell tolerance. Protein tyrosine phosphatase non receptor type 22 (PTPN22) is an important negative regulator of signal transduction from the T cell receptor. A single nucleotide polymorphism 1858C>T in the PTPN22 gene has been associated with various autoimmune disorders. We hypothesize that the PTPN22 mutation 1858C>T would be present in a higher proportion of ITP patients and patients who carry the mutation would have an increased concentration of autologous antiplatelet IgG on their platelet surface. We genotyped 45 patients with ITP by a PCR based restriction length polymorphism(RFLP) to identify the mutation 1858C>T in the gene PTPN22. Ten patients (22%) were heterozygous for the PTPN22 mutation and 2 were homozygous for the mutation (4.4%). This compares with a frequency of 8.6% observed in a published population study of 960 controls who did not have autoimmune disease (p<0.0001) (Am. J. Hu. Genetics2004:75:330). All of the 12 ITP patients who carried the PTPN22 mutation had an increase in autologous platelet surface IgG (≥1250 molecules IgG/platelet)(Schwartz Am. J. Heme1990;33:167). We conclude that the PTPN22 1858C>T mutation is present in an increased proportion of ITP patients and that all 12 of the PTPN22 mutation positive patients were also positive for autologous antiplatelet IgG. Our results indicate that PTPN22 increases the probability of developing ITP.