Objective:To perform whole-genome sequencing and phylogenetic analysis of the local endemic strain of Rodent Torque teno virus (RoTTV), RoTTV3-HMU1, found in Rattus norvegicus, Haikou City, Hainan Province, and establ...Objective:To perform whole-genome sequencing and phylogenetic analysis of the local endemic strain of Rodent Torque teno virus (RoTTV), RoTTV3-HMU1, found in Rattus norvegicus, Haikou City, Hainan Province, and establish a SYBR Green I based real-time PCR detection assay for RoTTV3.Methods: Based on the high-throughput genome sequencing analysis, specific primers were designed and the whole genome sequence was amplified by PCR and Sanger sequencing. Specific detection primers were designed based on the conserved sequences of RoTTV3. The recombinant plasmid contained the whole genome of RoTTV3-HMU1 was constructed as a standard control. The experimental conditions were optimized and the real-time PCR detection assay of RoTTV3 was established.Results: The genomic sequence of RoTTV carried by Rattus norvegicus in Haikou City was successfully sequenced. Phylogenetic analysis indicated that the virus belongs to the RoTTV3 genotype. In this experiment, the real-time PCR detection method of RoTTV3 was established. The standard curve generated had a wide dynamic range from 1×(102-108) copies/μL, with a linear correlation (R2=1.000). The melting curve analysis using SYBR Green showed only one specific melting peak and no primer-dimmers represented. The detection limit was 100 copies/reaction.Discussion: This study was the first report of the RoTTV in Hainan Islands, and its phylogenetic analysis was of great significance to the origin and evolution of RoTTV. The RoTTV3 real-time PCR detection method established in this experiment has a high sensitivity and good specificity, which lays a technical foundation for the epidemiological investigation of RoTTV3.展开更多
Background:Large-for-gestational age(LGA)newborns can increase the risk of metabolic syndrome.Previous studies have shown that the levels of maternal blood lipids,connecting peptide(C-peptide),insulin and glycosylated...Background:Large-for-gestational age(LGA)newborns can increase the risk of metabolic syndrome.Previous studies have shown that the levels of maternal blood lipids,connecting peptide(C-peptide),insulin and glycosylated hemoglobin(HbA_(1c))were significantly different between LGA and appropriate-for-gestational age(AGA)newborns.This study aimed to determine the effect of the levels of maternal lipids,C-peptide,insulin,and HbA_(1c) during late pregnancy on LGA newborns.Methods:This study comprised 2790 non-diabetic women in late pregnancy.Among their newborns,2236(80.1%)newborns were AGA,and 554(19.9%)newborns were LGA.Maternal and neonatal characteristics were obtained from questionnaires and their case records.The levels of maternal fasting serum apolipoprotein A1(ApoA1),triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),C-peptide,insulin and blood HbA_(1c) were measured.The chi-square and Mann-Whitney U test were used to analyze categorical variables and continuous variables between the AGA and LGA groups,respectively.Binary logistic regression analysis was made to determine the independent risk factors for LGA newborns.Results:Maternal TG,C-peptide,insulin and HbA_(1c) levels were signifi cantly higher in the LGA group than in the AGA group(P<0.05).The LGA group had signifi cantly lower levels of maternal TC,HDL-C and LDL-C than the AGA group(P<0.05).After adjustment for confounding variables,including maternal age,pre-pregnancy body mass index,education,smoking,annual household income,amniotic fluid volume,gestational hypertension,newborn gender and gestational age at blood collection,high maternal TG levels remained signifi cantly associated with LGA newborns(P<0.05).Conclusion:High maternal TG level during late pregnancy is signifi cantly associated with LGA newborns.展开更多
基金National natural science foundation of China,No.81860367,31460017,81672072Key Research and Development Program of Hainan Province,No.ZDYF2017091+1 种基金Special Project for Scientific Research of Institutions of Higher Learning in Hainan Province,No.Hnky2017ZD-16,Hnkyzx2014-08Open Fund Project of State key Laboratory of Virology in 2018.Project,No.2018IOV002.
文摘Objective:To perform whole-genome sequencing and phylogenetic analysis of the local endemic strain of Rodent Torque teno virus (RoTTV), RoTTV3-HMU1, found in Rattus norvegicus, Haikou City, Hainan Province, and establish a SYBR Green I based real-time PCR detection assay for RoTTV3.Methods: Based on the high-throughput genome sequencing analysis, specific primers were designed and the whole genome sequence was amplified by PCR and Sanger sequencing. Specific detection primers were designed based on the conserved sequences of RoTTV3. The recombinant plasmid contained the whole genome of RoTTV3-HMU1 was constructed as a standard control. The experimental conditions were optimized and the real-time PCR detection assay of RoTTV3 was established.Results: The genomic sequence of RoTTV carried by Rattus norvegicus in Haikou City was successfully sequenced. Phylogenetic analysis indicated that the virus belongs to the RoTTV3 genotype. In this experiment, the real-time PCR detection method of RoTTV3 was established. The standard curve generated had a wide dynamic range from 1×(102-108) copies/μL, with a linear correlation (R2=1.000). The melting curve analysis using SYBR Green showed only one specific melting peak and no primer-dimmers represented. The detection limit was 100 copies/reaction.Discussion: This study was the first report of the RoTTV in Hainan Islands, and its phylogenetic analysis was of great significance to the origin and evolution of RoTTV. The RoTTV3 real-time PCR detection method established in this experiment has a high sensitivity and good specificity, which lays a technical foundation for the epidemiological investigation of RoTTV3.
基金supported by grants from the"11th Five-Year Plan"and the"12th Five-Year Plan"from the National Science and Technology Issues Research,China(2009BAI80B03,2012BAI02B03)the Innovation Program for Early Screening and Intervention of Birth Defects,Zhejiang Province(2010R50045)the National Key Scientifi c Research Projects of China(973 Program)(2012CB944900).
文摘Background:Large-for-gestational age(LGA)newborns can increase the risk of metabolic syndrome.Previous studies have shown that the levels of maternal blood lipids,connecting peptide(C-peptide),insulin and glycosylated hemoglobin(HbA_(1c))were significantly different between LGA and appropriate-for-gestational age(AGA)newborns.This study aimed to determine the effect of the levels of maternal lipids,C-peptide,insulin,and HbA_(1c) during late pregnancy on LGA newborns.Methods:This study comprised 2790 non-diabetic women in late pregnancy.Among their newborns,2236(80.1%)newborns were AGA,and 554(19.9%)newborns were LGA.Maternal and neonatal characteristics were obtained from questionnaires and their case records.The levels of maternal fasting serum apolipoprotein A1(ApoA1),triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),C-peptide,insulin and blood HbA_(1c) were measured.The chi-square and Mann-Whitney U test were used to analyze categorical variables and continuous variables between the AGA and LGA groups,respectively.Binary logistic regression analysis was made to determine the independent risk factors for LGA newborns.Results:Maternal TG,C-peptide,insulin and HbA_(1c) levels were signifi cantly higher in the LGA group than in the AGA group(P<0.05).The LGA group had signifi cantly lower levels of maternal TC,HDL-C and LDL-C than the AGA group(P<0.05).After adjustment for confounding variables,including maternal age,pre-pregnancy body mass index,education,smoking,annual household income,amniotic fluid volume,gestational hypertension,newborn gender and gestational age at blood collection,high maternal TG levels remained signifi cantly associated with LGA newborns(P<0.05).Conclusion:High maternal TG level during late pregnancy is signifi cantly associated with LGA newborns.
