TaqMan Polymerase Chain Reaction Method Research Articles

Development of a TaqMan polymerase chain reaction detection method for the precise identification and quantification of an attenuated Eimeria maxima vaccine strain in poultry.

Avian coccidiosis, a parasitic disease prevalent in poultry, is caused by Eimeria species and leads to significant economic losses. The use of attenuated live oocyst vaccines has been adopted as an alternative to the use of anticoccidial drugs. However, the accurate detection and differentiation of vaccine strains from virulent ones remain challenging. Therefore, this study presents a novel TaqMan polymerase chain reaction (PCR) detection method that offers enhanced sensitivity, specificity, and reproducibility compared with traditional PCR techniques. Through whole-genome resequencing and bioinformatics analysis, we identified a molecular marker gene, Em_marker6, with a unique 21-base pair deletion specific to the Eimeria maxima attenuated vaccine strain. Optimized primers and probes targeting this marker enabled rapid quantification cycle value achievement and high fluorescence intensity. The standard curve's slope of -3.540 and correlation coefficient of 0.9971 confirmed precise quantification capabilities. The TaqMan PCR method detected as few as 30 plasmid DNA copies and 50 oocysts per reaction, outperforming traditional PCR techniques by an order of magnitude. No cross-reactivity was observed with other E. maxima wide-type strains or common intestinal pathogens, ensuring the exclusive detection of the E. maxima EMPY vaccine strain. Weekly testing over 3 weeks demonstrated minimal variability, indicating robust consistency in the method's application. Testing on 61 clinical samples revealed a 57.38% positivity rate for E. maxima species and 13.11% for the vaccine strain. The Em_marker6 gene exhibited genetic stability across multiple generations, confirming the detection method's robust stability for the attenuated E. maxima vaccine strain. This study significantly advances the field of avian coccidiosis research and control by providing a valuable tool for monitoring vaccine purity and preventing inadvertent infections in vaccinated flocks, aligning with global efforts to curb antibiotic use in animal feed.

Association of hyaluronan and proteoglycan link protein 1 gene with the need of home oxygen therapy in premature Japanese infants with bronchopulmonary dysplasia

Background Bronchopulmonary dysplasia (BPD) has a lasting effect on the respiratory function of infants, imposing chronic health burdens. BPD is influenced by various prenatal, postnatal, and genetic factors. This study explored the connection between BPD and home oxygen therapy (HOT), and then we examined the association between HOT and a specific single-nucleotide polymorphism (SNP) in the hyaluronan and proteoglycan link protein 1 (HAPLN1) gene among premature Japanese infants. Materials and methods Prenatal and postnatal data from 212 premature infants were collected and analyzed by four SNPs (rs975563, rs10942332, rs179851, and rs4703570) around HAPLN1 using the TaqMan polymerase chain reaction method. The clinical characteristics and genotype frequencies of HAPLN1 were assessed and compared between HOT and non-HOT groups. Results Individuals with AA/AC genotypes in the rs4703570 SNP exhibited significantly higher HOT rates at discharge than those with CC homozygotes (odds ratio, 1.20, 95% confidence interval, 1.07–1.35, p = .038). A logistic regression analysis determined that CC homozygotes in the rs4703570 SNP did not show a statistically significant independent association with HOT at discharge. Conclusions Although our study did not reveal a correlation between HAPLN1 and the onset of BPD, we observed that individuals with CC homozygosity at the rs4703570 SNP exhibit a reduced risk of HOT.

Analysis of two single-nucleotide polymorphisms (rs2241766 and rs1501299) of the adiponectin gene in patients with coronary artery disease and coronary slow flow.

Aim To investigate the association of two single-nucleotide polymorphisms (SNPs) of the adiponectin gene (+45 T>G [rs2241766] and +276 G>T [rs1501299]) in patients with coronary artery disease (CAD), coronary slow flow (CSF) and in healthy subjects. Methods The study investigated 90 patients: 30 patients with coronary artery disease (the CAD group), 30 with only coronary slow flow (the CSF group) and 30 healthy patients. Genotyping was carried out for two of these SNPs in the adiponectin gene using the TaqMan polymerase chain reaction (PCR) method. Results There were no significant differences in the frequencies of polymorphism +45T>G (rs2241766) genotype (p=0.464) and the allele (p=0.362). There were also no significant differences between the +276 G>T (rs1501299) genotype and the allele (p=0.624 and p=0.281, respectively). Conclusion Our study indicated that in patients with CAD, CSF, and in healthy subjects, the SNP +276G>T and +45T>G alleles of the adiponectin gene were not associated.

Association of SOCS3 gene polymorphisms with insulin resistance in Xinjiang Uygur population

To assess the association of suppressor of cytokine signaling 3 (SOCS3) gene polymorphisms with insulin resistance (IR) in Xinjiang Uygur population. A cross-sectional study on metabolic diseases, e.g., obesity, was conducted on ethnic Uygurs in Hetian, Xinjiang of China. A total of 1292 Uygur individuals were enrolled. The sample size for IR subjects [homeostasis model assessment for insulin resistance (HOMA-IR) ≥ 2.96] was 323, whereas that for non-IR controls was 969 (HOMA-IR < 2.96). Representative variations were selected from database and genotyped with TaqMan polymerase chain reaction method. For the relatively isolated population from a homogeneous environment, a case-control study was conducted to assess the association between variations of SOCS3 gene and IR. A significant difference in genotype distribution of rs4969168 was detected between IR and control groups in males (chi square =7.216, P=0.027). Although the IR-related quantitative phenotypes did not significantly differ between individuals with GG, AG and AA genotypes of rs4969168 in the overall, male and female population (P > 0.05), the mean of body mass index and the median of fasting insulin increased in individuals with GG, AG, AA genotypes in males. Haplotype 2 (rs12953258C-rs4969168A-rs9914220C) was significantly associated with a higher prevalence of IR in males (P=0.023). Logistic regression analysis indicated that AG genotype of rs4969168 variation is a protective factor for IR in males (OR=1.772, 95% confidence interval: 1.081-2.906, P=0.023). Our study suggested that the rs4969168 polymorphism of SOCS3 gene is associated with IR in ethnic Uygur population from Xinjiang, China.

Genetic variations of the ADIPOQgene and risk of prostate cancer in Chinese Han men.

Adiponectin secreted by adipose tissue has been implicated in prostate carcinogenesis. Genetic variations in ADIPOQ are thought to influence the activity of adiponectin, thus relating to cancer occurrence. In this hospital-based case-control study of 917 prostate cancer (PCa) cases and 1036 cancer-free controls, we evaluated the association of single nucleotide polymorphisms in ADIPOQ with risk of PCa and adiponectin levels in Chinese Han men. Variants of ADIPOQ were genotyped by Taqman polymerase chain reaction method. The plasma adiponectin concentrations were measured by enzyme-linked immunosorbent assay (ELISA) in a subset of cases and controls. We found that the ADIPOQ rs3774262 variant AA genotype was associated with both decreased PCa risk [adjusted odds ratio (OR): 0.66, 95% confidence interval (CI) =0.48–0.92] and increased plasma adiponectin levels (P = 0.036 and 0.043), with significant difference by tumor grade, clinical stage, and aggressiveness. A significant interaction between ADIPOQ rs3774262 and body mass index was observed in modifying the risk of PCa (P = 6.7 × 10−3). ADIPOQ rs266729 and rs182052 were not related to PCa risk or plasma adiponectin levels. Our data support that ADIPOQ rs3774262 may affect PCa risk in combination with plasma adiponectin levels in Chinese Han men. It may contribute to the molecular basis for the association between obesity and PCa.

GW24-e2291 Association of SOCS3 genetic polymorphisms with insulin resistance in Xinjiang Uygur population

ObjectivesTo investigate the association between suppressor of cytokine signalling 3(SOCS3) genetic polymorphisms and insulin resistance (IR) in Xinjiang Uygur population.MethodsIn this cross-sectional study on the metabolic diseases (e.g. obesity) among...

GW24-e2293 Association Study of the GIRK4 Polymorphisms and Essential Hypertension in Obesity Uygur of Southern Xinjiang

ObjectivesTo analyse the association between the G protein-coupled inwardly-rectifying potassium channels4 (GIRK4) gene and Essential hypertension in Uygur in Xinjiang Uygur Autonomous Region.MethodsA total of 1194 (male 461, female733) Uygur...

Common variants in adiponectin gene are associated with coronary artery disease and angiographical severity of coronary atherosclerosis in type 2 diabetes

BackgroundAdiponectin, an adipokine facilitating insulin action, has antiatherogenic effects. This study investigated whether common single nucleotide polymorphisms (SNPs) in the adiponectin gene influenced plasma adiponectin level and whether they were associated with the risk of coronary artery disease (CAD) and its angiographical severity in type 2 diabetes in Chinese population.Methods11 tagging SNPs were genotyped in 1110 subjects with or without CAD in type 2 diabetes. Variants of adiponectin gene were determined by Taqman polymerase chain reaction method. The plasma adiponectin concentrations were measured by sandwich enzyme-linked immunosorbent assay. The severity and extent of coronary atherosclerosis were assessed using the angiographic Gensini score and Sullivan Extent score.ResultsAmong the 11 SNPs, the minor G allele of SNP rs266729 was significantly associated with higher odds of CAD (odds ratio (95% CI) = 1.49 (1.10 - 2.16), P = 0.022) after adjusting for covariates. In stepwise multivariate logistic regression, SNP rs266729 was a significant independent factor of CAD. Multivariate linear regression analysis revealed that rs266729 (β = −0.101, P < 0.0001), rs182052 (β = −0.044, P = 0.0035), and rs1501299 (β = 0.073, P < 0.0001) were significantly associated with adiponectin level, and also indicated that the minor G allele of SNP rs266729 had higher Gensini score (β = 0.139, P < 0.001) and Sullivan Extent score (β = 0.107, P < 0.001). Haplotypes analysis revealed different haplotype distributions in case and control subjects (P = 0.0003), with two common haplotypes GGG and GAG of the rs266729, rs182052, and rs1501299 being associated in heterozygotes with a greater than threefold increase in cardiovascular risk (odds ratio (95% CI)=3.39 (1.83 - 6.30), P = 0.0001).ConclusionsIn our population, genetic variants in the adiponectin gene influence plasma adiponectin levels, and one of them is a strong determinant of CAD susceptibility and its angiographical severity in type 2 diabetes. This study has provided further evidence for a role of adiponectin in the development of CAD.

Real-time TaqMan polymerase chain reaction–based genus-identification and pyrosequencing-based species identification of Campylobacter jejuni, C. coli, C. lari, C. upsaliensis, and C. fetus directly on stool samples

A new method was developed for Campylobacter identification and applied directly on 599 stool samples from diarrhoeagenic patients. Here, the gyrase B gene of Campylobacter was targeted in a 2-step process: first, TaqMan polymerase chain reaction (PCR)–based identification of C. jejuni, C. coli, C. upsaliensis, C. lari, and C. fetus at the genus level, and, second, pyrosequencing-based identification at the species level. The TaqMan PCR method was compared to culturing and identified 87 Campylobacter-positive samples of which 64 were culture positive. Among the discrepant 23 samples, 18 were confirmed positive by conventional PCR, underlining a significant increase in diagnostic yield by use of this molecular and culture-independent method. For species identification, the pyrosequencing method was compared to conventional PCR and among the 87 TaqMan PCR-positive samples, 74 Campylobacter species were identified by both methods, 10 samples gave discrepant results, and 3 samples were negative by both methods.

Identification of Francisella tularensis by both culture and real-time TaqMan PCR methods from environmental water specimens in outbreak areas where tularemia cases were not previously reported

Tularemia is an endemic disease in Turkey. In this study, we aimed to detect Francisella tularensis by two methods from natural water supplies thought to cause tularemia epidemiologically. A total of 154 water specimens from three different outbreaks caused by drinking water were collected. Water specimens were cultured on antibiotic-added cysteine heart agar base with blood and incubated at 37°C in a humidified atmosphere containing 5% CO(2) for 4-10days. The suspected colonies were confirmed by F. tularensis antiserum (BD) and the real-time TaqMan polymerase chain reaction (PCR) method. DNA was isolated from samples obtained from filters. The primer and probe sets targeting the ISFtu2 genome were used. A total of four F. tularensis isolates were obtained from 154 water samples. At the same time, the presence of F. tularensis DNA from 17 water specimens was shown by the real-time TaqMan PCR method. Although the DNA presence of F. tularensis has been detected from water sources by the PCR method in Turkey up to now, there has been no isolation directly from water specimens by culture. In this study, the determination of F. tularensis from water sources has been exhibited as the first data by both culture and real-time TaqMan PCR methods.

Investigation of Ureaplasma urealyticum biovars and their relationship with antimicrobial resistance

Purpose: To develop Taqman fluorescence quantitative polymerase chain reaction (PCR) method for investigating the characteristics of the distributions of Ureaplasma urealyticum (UU) biovars and to explore the relationship between UU biovars and antimicrobial resistance. Materials and Methods: By the method of culture, Ureaplasma species were detected. Taqman fluorescence quantitative PCR for detecting UU biovars were developed and UU clinical isolates were detected to distinguish biovars. The broth micro-dilution susceptibility testing methods were used to determine UU susceptibility. Results: By Taqman PCR method, UU biovars was successfully detected. Of 126 samples, biovar 1 was found in 73 (57.94%). There was a statistical difference between genital-urinary tract infection group and asymptomatic group (P<0.05). In the region, UU biovar 1 to 9 kinds of agents kept higher susceptibility rates, but biovar 2 maintained higher susceptibility rates only to tetracyclines. Compared with biovar 1, UU biovar 2 resistance rates to 7 kinds of agents were higher (P<0.05). Conclusions: (1) Our new established Taqman PCR method is a useful tool for screening UU biovars. (2) UU biovar 1 predominated in asymptomatic population; whereas in genital-urinary tract infection population UU biovar 2 had a higher proportion. (3) The characteristics of drug resistance were different between UU biovars. Overall, both two biovars remained higher susceptibility rates to tetracyclines. A majority of biovor 1 strains were sensitive to macrolides and quinolones; while only a small number of biovar 2 strains kept sensitive to roxithromycin and quinolones, a large proportion of biovar 2 strains were found in intermediate ranges.

Association of the genetic variations of bone morphogenetic protein 7 gene with diabetes and insulin resistance in Xinjiang Uygur population

To investigate the association between the genetic variations of the functional region in bone morphogenetic protein gene (BMP7) with type 2 diabetes mellitus in Chinese Uygur individuals. A case-control study was conducted based on epidemiological investigation. A total of 717 Uygur subjects (276 males and 441 females) were selected and divided into two groups: diabetes mellitus group (n = 502, 191 males and 311 females) and control group (n = 215, 85 males and 130 females). All exons, flanking introns and the promoter regions of (BMP7) gene were sequenced in 48 Uygur diabetics. Representative variations were selected according to the minor allele frequency (MAF) and linkage disequilibrium and genotyped using the TaqMan polymerase chain reaction method in 717 Uygur individuals, a relatively isolated general population in a relatively homogeneous environment and a case-control study was conducted to test the association between the genetic variations of (BMP7) gene and type 2 diabetes mellitus. Five novel and 8 known variations in the (BMP7) gene were identified. All genotype distributions were tested for deviations from Hardy-Weinberg equilibrium (P> 0.05). There was significant difference of genotype distribution of rs6025422 between type 2 diabetes mellitus and control groups in the male population (P< 0.05, P adjusted > 0.05), but there was no difference in total and female population (P> 0.05). And the means of fasting blood glucose (FBG), fasting insulin and HOMA-index significantly decreased in individuals with AA, AG and GG genotypes of rs6025422 in male population (P< 0.05), but not in total and female population (P> 0.05). The logistic regression analysis showed that GG genotype of rs6025422 variation might be a protective factor for diabetes in male (OR= 0.637, 95% confidence interval 0.439-0.923, P< 0.05). The present study suggests that the rs6025422 polymorphism in (BMP7) gene may be associated with diabetes mellitus and insulin resistance in Uygur men.

Single-nucleotide polymorphism in the hyaluronan and proteoglycan link protein 1 (HAPLN1) gene is associated with spinal osteophyte formation and disc degeneration in Japanese women

Spinal osteoarthritis including disc degeneration is a very common condition in the axial skeletons of aged people. Recently, spinal osteoarthritis has been shown to be influenced by specific genetic risk factors. Vertebral osteophytes, endplate sclerosis, and intervertebral disc narrowing are recognized as radiographic features of spinal disc degeneration. HAPLN1 is a key component of the cartilage extracellular matrix; thus, variations in this gene may affect the pathogenesis of cartilage-related diseases such as spinal degeneration. Here, we examine the association between an HAPLN1 gene polymorphism and the radiographic features of spinal degeneration. We evaluated the degree of endplate sclerosis, osteophyte formation, and disc space narrowing in 622 Japanese postmenopausal women. Four SNPs in the HAPLN1 gene-in the 5' flanking region, intron 1, intron 2, and intron 4-were analyzed using the TaqMan polymerase chain reaction method. We found that compared to subjects with the CC or CT genotype, those with the TT genotype for an SNP at intron 2 (rs179851) were significantly overrepresented among the subjects with higher scores for osteophyte formation (P = 0.0001; odds ratio 2.12; 95% confidence interval 1.45-3.11, as determined by logistic regression analysis) and disc space narrowing (P = 0.0057; odds ratio 1.83; 95% confidence interval 1.19-2.83). Consistent with the involvement of the HAPLN1 gene in cartilage metabolism, a variation in a specific HAPLN1 gene locus may be associated with spinal degeneration.

Associations between genetic variations in the FURIN gene and hypertension

BackgroundHypertension is a complex disease influenced by multiple genetic and environmental factors. The Kazakh ethnic group is characterized by a relatively high prevalence of hypertension. Previous research indicates that the FURIN gene may play a pivotal role in the renin-angiotensin system and maintaining the sodium-electrolyte balance. Because these systems influence blood pressure regulation, we considered FURIN as a candidate gene for hypertension. The purpose of this study was to systematically investigate the association between genetic variations in the FURIN gene and essential hypertension in a Xinjiang Kazakh population.MethodsWe sequenced all exons and the promoter regions of the FURIN gene in 94 hypertensive individuals to identify genetic variations associated with the disorder. Genotyping was performed using the TaqMan polymerase chain reaction method for four representative common single nucleotide polymorphisms (SNPs, -7315C > T, 1970C > G, 5604C > G, 6262C > T) in 934 Kazakh Chinese people. One SNP (1970C > G) was replicated in 1,219 Uygur Chinese people.ResultsNine novel and seven known single nucleotide polymorphisms were identified in the FURIN gene. The results suggest that 1970C > G was associated with a hypertension phenotype in Kazakh Chinese (additive model, P = 0.091; dominant model, P = 0.031, allele model, P = 0.030), and after adjustment with logistic regression analysis, ORs were 1.451 (95%CI 1.106-1.905, P = 0.008) and 1.496 (95% 1.103-2.028, P = 0.01) in additive and dominant models, respectively. In addition, the association between 1970C > G and hypertension was replicated in Uygur subjects (additive model, P = 0.042; dominant model, P = 0.102; allele model, P = 0.027) after adjustment in additive and dominant models, ORs were 1.327 (95% 1.07-1.646), P = 0.01 and 1.307 (95%CI 1.015-1.681, P = 0.038), respectively. G allele carriers exhibited significant lower urinary Na+ excretion rate than non-carriers in the Kazakh Chinese population (152.45 ± 76.04 uM/min vs 173.33 ± 90.02 uM/min, P = 0.007).ConclusionOur results suggest that the FURIN gene may be a candidate gene involved in human hypertension, and that the G allele of 1970C > G may be a modest risk factor for hypertension in Xinjiang Kazakh and Uygur populations.

Single-nucleotide polymorphisms in SMAD3 are associated with chronic obstructive pulmonary disease

The aim of this study was to determine the frequencies of single-nucleotide polymorphisms (SNPs) in the Smad3 gene in the Chinese population and their possible association with chronic obstructive pulmonary disease (COPD). The frequency distribution of nine SNPs in the introns of the Smad3 gene was determined in both the COPD patients and control groups by the TaqMan polymerase chain reaction method using a minor groove binder probe. The genotype distribution of the rs28683050 polymorphism in control subjects was as follows: CC, 67.6%; CT, 27.0% and TT, 5.4%; and in COPD patients was as follows: CC, 47.0%; CT, 38.8% and TT, 14.2%. Therefore, a significant difference in allelic and genotypic frequencies between the COPD patients and control subjects is observed. Their genotypic distribution was not significantly different from that predicted by Hardy-Weinberg's equilibrium. The frequency of the TT genotype in the COPD patients was significantly higher than that in controls (14.2% versus 5.4%, odds ratio [OR] = 3.762, 95% confidence interval: 1.650-8.581, P = 0.002), and the frequency of the T allele in the COPD patients was significantly higher than that in controls (81.1% versus 66.4%, OR = 1.774, 95% confidence interval: 1.354-2.324, P = 0.001). These data show that the rs28683050 polymorphism of the Smad3 gene may be associated with COPD.

Association of genetic variations of prostasin gene with the essential hypertension in Xinjiang Kazakhs

To explore the relationship between genetic variations of prostasin gene and essential hypertension (EH) in Xinjiang Kazakhs. Totally 938 Fukang City residents who were older than 30 years were enrolled in this study using cluster random sampling method. Standardized questionnaire and physical examination were performed. Among them there 451 EH patients (EH group) and 478 normotensive (NT) subjects (NT group) according to Guidelines of Prevention and Control for Hypertension in 2005. All the exons and promoter regions of prostasin gene were sequenced in 94 EH patients. Representative variations (297A>C, 2827C>T, and E342K) were genotyped using TaqMan polymerase chain reaction method in all 938 subjects. The frequencies of genotypes were compared between the EH and NT groups. Ten variations were found as follows: -36G>C, -27C>T, 78G>A, 81G>C (rs8049043), 297A>C, 350C>T, 351A>C, 2827C>T, 3482G>A (E342K), and 3783A>G. E342K and 2827C>T were successfully genotyped. E342K mutation was identified in only one hypertensive patient. CC, CT, and TT genotypes existed in 2827C>T polymorphism. The frequencies of CC, CT, and TT were 81.0%, 17.3, and 1.7% in EH group and 80.3%, 18.9%, and 0.8% in NT group, respectively. The frequencies of C and T alleles were 89.6% and 10.4% in EH group and 89.8% and 10.2% in NT group, respectively. The distribution of genotypes and allele frequencies were not significantly different between these two groups (chi2=2.048, P=0.353 and chi2=0.001, P=0.973). Blood pressure was not significantly among subjects with these three genotypes (P>0.05). The EH of Xinjiang Kazakhs is not associated with 2827C>T polymorphism in the prostasin gene. E342K mutation in the prostasin gene may contribute partly to the hypertensive phenotype in this population.

Association of adiponectin genotype polymorphisms at positions 45 and 276 with obstructive sleep apnea hypopnea syndrome

Objective To investigate the relationship between adiponectin genotype polymorphisms and obstructive sleep apnea hypopnea syndrome (OSAHS). Methods Using the TaqMan polymerase chain reaction(PCR) method, the single nucleotide polymorphisms(SNP) at positions 45 and 276 in the adiponectin gene were determined in Chinese of the Han nationality in the Nanjing district. The OSAHS group consisted of 78 patients, and the control group contained 40 subjects. The association of adiponectin genotype polymorphisms at positions 45 and 276 with obstructive sleep apnea hypopnea syndrome was analyzed. Results No evidence of a direct association was found between OSAHS and adiponectin genotype SNP at positions 45 and 276( P > 0.05). However, compared with those OSAHS patients having G/T+T/T genotype at position 276, the OSAHS patients with the G/G genotype showed a greater neck circumference(NC), a prolonged duration of the longest apnea event, and elevated levels of blood cholesterol and low-density lipoprotein cholesterol( P < 0.05). Conclusion No direct association was detected between OSAHS and adiponectin genotype distribution at positions 45 and 276 in Chinese of Han nationality in the Nanjing district. However, OSAHS patients with the adiponectin G/G genotype at position 276 had a larger NC and the longest apnea event compared to those having the adiponectin SNP276 G/T +T/T genotype. This may have an indirect influence on the development of OSAHS,

Human uncoupling protein 2 and 3 genes are associated with obesity in Japanese

Human uncoupling proteins (UCPs) are mitochondrial proteins that are involved in the control of energy metabolism and the pathophysiology of obesity. Although there have been several reports on the association between the UCP2/UCP3 locus and the obesity, there have been no haplotype-based case-control studies with gender-specific analysis. The aim of this study was to examine whether there is an association between the UCP2/UCP3 locus and the obesity in the Japanese population when using a single nucleotide polymorphism (SNP)-based and haplotype-based case-control study with gender-specific analysis. We examined a group consisting of 551 subjects, of which 369 were non-obese and 182 were overweight and/or obese. We selected one nonsynonymous SNP (rs660339: Ala55Val) as a genetic marker. Genotyping for all subjects was performed by the TaqMan polymerase chain reaction (PCR) method. Although the overall distributions of genotype and allele were not significantly different between the non-obese and the obese groups, the overall distributions of the genotype were significantly different in men (P = 0.030). In the obese group, male subjects with the Val allele were significantly more frequent in both association studies. There was a significant difference in the overall distribution of the haplotype (UCP3 rs180049, UCP3 rs2075577, UCP2 rs660339) between the weight groups (P = 0.010), and in women, there was a significant difference (P = 0.042) in the overall distribution of the haplotype (UCP3 rs2075577, UCP2 rs660339). Nonsynonymous rs660339 in the human UCP2 gene in men, and the haplotype (UCP3 rs2075577-UCP2 rs660339) in women might be good obesity markers.

Association of a Single Nucleotide Polymorphism in the Insulin-Like Growth Factor-1 Receptor Gene With Spinal Disc Degeneration in Postmenopausal Japanese Women

An association study investigating the genetic etiology for spinal disc degeneration. To determine the association of single-nucleotide polymorphism (SNP) in the insulin-like growth factor-1 receptor (IGF1R) with spinal disc degeneration. Insulin-like growth factor-1 (IGF-1) signaling pathway is involved in cartilage development and homeostasis, suggesting that genetic variations of genes involved in this pathway may affect the pathogenesis of cartilage-related diseases, such as disc degeneration. We evaluated the presence of endplate sclerosis, osteophytes, and narrowing of disc spaces in 434 Japanese postmenopausal women. A SNP in the IGF1R gene at intron 1 was determined using TaqMan polymerase chain reaction method. We compared those who carried the G allele (GG or GC, n = 290) with those who did not (CC, n = 144). We found that the subjects with the G allele (GG or GC) were significantly over-represented in the subjects having higher disc narrowing score (P = 0.0033; odds ratio, 2.04; 95% confidence interval, 1.27-3.29 by logistic regression analysis). We suggest that a genetic variation at the IGF1R gene locus is associated with spinal disc degeneration, in line with the involvement of the IGF1R gene in the cartilage metabolism.

Abstract 2911: T-182C Polymorphism in the Norepinephrine Transporter Gene (SLC6A2) and Risk of Cardiovascular Events in Patients with Dilated Cardiomyopathy: Association of the T allele with a Reduced Risk for Cardiovascular Events

Backgrounds: The increase in sympathetic activity is one of the hallmarks of the chronic heart failure and has been shown to have an important impact on survival in patients with dilated cardiomyopathy (DCM). Norepinephrine transporter (NET) recaptures as much as 90% of released norepinephrine in the heart, making it a critical mediator of norepinephrine inactivation and presynaptic catecholamine homeostasis. Recently, the polymorphism of NET T-182C was reported to be associated with the improvement of left ventricular systolic function by beta-blockers in patients with DCM. However, the association between the incidence of cardiovascular events and the NET polymorphism in patients DCM has not been understood. The purpose of this study was to evaluate the effect of this polymorphism on the incidence of cardiovascular events in patients with DCM. Methods: Eighty-three genetically unrelated patients with nonfamilial DCM (64 males, mean age at initial clinical evaluation 59 ± 14 years) were enrolled in this study. An adverse cardiovascular event was defined as cardiac death or hospitalization for cardiac reasons. The time to the first adverse cardiovascular event was analyzed by the Kaplan-Meier method. The TaqMan polymerase chain reaction method was used for the determination of genotypes of the NET T-182C gene (SLC6A2) (rs#2242446). Results: The distribution of the NET T-182C genotypes (T/T, T/C, and C/C) was 43%, 45%, and 12%, respectively. The NET T-182C T allele frequency was 0.63. During a mean follow-up period of 45 months, 20 cardiovascular events had occurred. Eight patients died from cardiac cause (1 from pump failure and 7 from sudden cardiac death) and there were 12 hospitalizations for new onset or worsening of heart failure symptoms. The cardiovascular event rate of the patients carrying the T allele (T/T and T/C genotype, n = 73) was significantly lower than that of the patients not carrying the T allele (C/C genotype, n = 10) (p = 0.03). Conclusion: Our data suggested that the T allele of the NET T-182C gene may be a protective factor against cardiovascular events in patients with DCM.