200例CDHR3基因单核苷酸多态性与人鼻病毒所致婴幼儿下呼吸道感染的关系
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摘要
目的探讨CDHR3基因单核苷酸多态性与人鼻病毒(human rhinoviruses,HRV)所致儿童急性下呼吸道感染的关系。方法收集2012年1月至2014年11月因HRV所致急性下呼吸道感染的患儿鼻咽抽吸物,提取RNA逆转录后,采用PCR方法扩增CDHR3基因片段,测序后进行rs6967330位点基因分型。利用PCR的方法扩增HRV的VP4基因进行测序分型,实时荧光定量PCR方法进行HRV载量测定,并回顾性分析rs6967330位点与患儿临床特征的相关性。结果共纳入200例患儿,其中AA/AG基因型30例(15. 0%),GG基因型170例(85. 0%)。HRV所致急性下呼吸道感染患儿中,AA/AG基因型与GG基因型相比更容易出现喘息症状(86. 7%vs 57. 1%,P=0. 002),而且更易致重症肺炎(30. 0%vs 11. 2%,P=0. 018),一级亲属也更多具有特应性疾病病史(16. 7%vs 5. 88%,P=0. 039)。Logistic回归分析发现rs6967330 A是HRV感染患儿喘息(OR:4. 585,95%CI:1. 446～14. 541,P=0. 010)及发展为重症病例(OR:3. 379,95%CI:1. 009～11. 315,P=0. 048)的危险因素。而HRV病毒亚型及载量在不同基因型间差异没有统计学意义。结论 CDHR3基因单核苷酸多态性位点rs6967330可能与HRV感染后的重症及喘息相关。
Objective To explore the association of single nucleotide polymorphism( SNP) at rs6967330 of cadherin-associated family member 3( CDHR3) gene with acute respiratory tract infection caused by human rhinoviruses( HRVs) in children. Methods From January,2012 to November,2014,we collected specimens of nasopharyngeal aspirates from 200 children with acute lower respiratory tract infection caused by HRVs. CDHR3 gene fragment was amplified by PCR from the samples and sequenced for rs6967330 genotyping. The VP4 gene of HRV was amplified with PCR for subsequent sequencing,and realtime fluorescent quantitative PCR was used to estimate the virus load. The correlation between rs6967330 genotypes and the clinical features of the children was analyzed. Results Of the 200 children with acute lower respiratory tract infection caused by HRVs,30( 15. 0%) had AA/AG genotype and 170( 85. 0%) had GG genotype of rs6967330. The children with AA/AG genotype were more likely to have wheezing symptoms than those with GG genotype( 86. 7% vs 57. 1%,P = 0. 002),and were also more susceptible to severe pneumonia( 30. 0% vs 11. 2%,P = 0. 018); the first-degree relatives of the children with AA/AG genotype were also more likely to have a history of atopic diseases( 16. 7% vs 5. 88%,P = 0. 039). Logistic regression analysis revealed that rs6967330 A was a risk factor for wheezing in children with HRV infection( OR: 4. 585,95% CI: 1. 446 ～ 14. 541,P = 0. 010) and also for progression into severe cases( OR: 3. 379,95% CI:1. 009 ～ 11. 315,P = 0. 048). No significant differences in HRV subtypes or viral loads were found between the children with different genotypes of rs6967330. Conclusion The SNP of CDHR3 gene at rs6967330 may be associated with severe cases and wheezing after lower respiratory tract infection with HRV in children.
Objective To explore the association of single nucleotide polymorphism( SNP) at rs6967330 of cadherin-associated family member 3( CDHR3) gene with acute respiratory tract infection caused by human rhinoviruses( HRVs) in children. Methods From January,2012 to November,2014,we collected specimens of nasopharyngeal aspirates from 200 children with acute lower respiratory tract infection caused by HRVs. CDHR3 gene fragment was amplified by PCR from the samples and sequenced for rs6967330 genotyping. The VP4 gene of HRV was amplified with PCR for subsequent sequencing,and realtime fluorescent quantitative PCR was used to estimate the virus load. The correlation between rs6967330 genotypes and the clinical features of the children was analyzed. Results Of the 200 children with acute lower respiratory tract infection caused by HRVs,30( 15. 0%) had AA/AG genotype and 170( 85. 0%) had GG genotype of rs6967330. The children with AA/AG genotype were more likely to have wheezing symptoms than those with GG genotype( 86. 7% vs 57. 1%,P = 0. 002),and were also more susceptible to severe pneumonia( 30. 0% vs 11. 2%,P = 0. 018); the first-degree relatives of the children with AA/AG genotype were also more likely to have a history of atopic diseases( 16. 7% vs 5. 88%,P = 0. 039). Logistic regression analysis revealed that rs6967330 A was a risk factor for wheezing in children with HRV infection( OR: 4. 585,95% CI: 1. 446 ～ 14. 541,P = 0. 010) and also for progression into severe cases( OR: 3. 379,95% CI:1. 009 ～ 11. 315,P = 0. 048). No significant differences in HRV subtypes or viral loads were found between the children with different genotypes of rs6967330. Conclusion The SNP of CDHR3 gene at rs6967330 may be associated with severe cases and wheezing after lower respiratory tract infection with HRV in children.
引文
[1] PALMENBERG A C. Rhinovirus C,asthma,and cell surface expression of virus receptor CDHR3[J]. J Virol,2017,91(7):e00072-C00017. DOI:10. 1128/JVI. 00072-17.
[2] JACOBS S E,LAMSON D M,ST GEORGE K,et al. Human rhinoviruses[J]. Clin Microbiol Rev,2013,26(1):135-162. DOI:10. 1128/CMR. 00077-12.
[3]郑首燕.支气管哮喘急性发作患儿鼻病毒检出率及其分子流行病学研究[D].重庆:重庆医科大学,2016.ZHENG S Y. Detection rate of rhinovirus in children with acute asthma attack and its molecular epidemiology[D].Chongqing:Chongqing Medical University,2016.
[4] KANAZAW A J,MASUKO H,YATAGAI Y,et al. Genetic association of the functional CDHR3 genotype with early-onset adult asthma in Japanese populations[J]. Allergol Int,2017,66(4):563-567. DOI:10. 1016/j. alit. 2017. 02. 012.
[5] BOCHKOV Y A,WATTERS K,ASHRAF S,et al. Cadherin-related family member 3,a childhood asthma susceptibility gene product,mediates rhinovirus C binding and replication[J]. Proc Natl Acad Sci USA,2015,112(17):5485-5490. DOI:10. 1073/pnas. 1421178112.
[6] HEIM A,EBNET C,HARSTE G,et al. Rapid and quantitative detection of human adenovirus DNA by real-time PCR[J]. J Med Virol,2003,70(2):228-239. DOI:10. 1002/jmv. 10382.
[7] ALLANDER T,JARTTI T,GUPTA S,et al. Human bocavirus and acute wheezing in children[J]. Clin Infect Dis,2007,44(7):904-910. DOI:10.1086/512196.
[8] COIRAS M T,AGUILAR J C,GARC A M L,et al. Simultaneous detection of fourteen respiratory viruses in clinical specimens by two multiplex reverse transcription nested-PCR assays[J]. J Med Virol,2004,72(3):484-495. DOI:10.1002/jmv. 20008.
[9] COIRAS M T,P REZ-BRE A P,GARC A M L,et al. Simultaneous detection of influenza A,B,and C viruses,respiratory syncytial virus,and adenoviruses in clinical samples by multiplex reverse transcription nested-PCR assay[J]. J Med Virol,2003,69(1):132-144. DOI:10. 1002/jmv. 10255.
[10]肖秋艳.重庆地区人鼻病毒和肠道病毒D68在呼吸道感染住院患儿中的分子流行病学研究[D].重庆:重庆医科大学,2015.XIAO Q Y. Epidemiological analysis of human rhinovirus and enterovirus in hospitalized children with respiratory infections in Chongqing[D]. Chongqing:Chongqing Medical University,2015.
[11] CHANG E H,WILLIS A L,MCCRARY H C,et al. Association between the CDHR3 rs6967330 risk allele and chronic rhinosinusitis[J]. J Allergy Clin Immunol,2017,139(6):1990-1992. e2. DOI:10. 1016/j. jaci. 2016. 10. 027.
[12] ZHAO B,YU X,WANG C,et al. High human bocavirus viral load is associated with disease severity in children under five years of age[J]. PLo S ONE,2013,8(4):e62318. DOI:10. 1371/journal. pone. 0062318.
[13] B NNELYKKE K,SLEIMAN P,NIELSEN K,et al. A genome-wide association study identifies CDHR3 as a susceptibility locus for early childhood asthma with severe exacerbations[J]. Nat Genet,2014,46(1):51-55. DOI:10. 1038/ng. 2830.
[14] BAI J,SMOCK S L,JACKSON G R Jr,et al. Phenotypic responses of differentiated asthmatic human airway epithelial cultures to rhinovirus[J]. PLo S ONE,2015,10(2):e0118286. DOI:10. 1371/journal. pone. 0118286.
[15] STENBERG HAMMAR K, NIESPODZIANA K, VAN HAGE M,et al. Reduced CDHR3 expression in children wheezing with rhinovirus[J]. Pediatr Allergy Immunol,2018,29(2):200-206. DOI:10. 1111/pai. 12858.
[16] HUSBY A,PASANEN A,WAAGE J,et al. CDHR3 gene variation and childhood bronchiolitis[J]. J Allergy Clin Immunol,2017,140(5):1469-1471. e7. DOI:10. 1016/j.jaci. 2017. 06. 044.
[17] LAUINGER I L,BIBLE J M,HALLIGAN E P,et al. Patient characteristics and severity of human rhinovirus infections in children[J]. J Clin Virol,2013,58(1):216-220. DOI:10. 1016/j. jcv. 2013. 06. 042
[18] LINDER J E,KRAFT D C,MOHAMED Y,et al. Human rhinovirus C:age,season,and lower respiratory illness over the past 3 decades[J]. J Allergy Clin Immunol,2013,131(1):69-77. e1-6. DOI:10. 1016/j. jaci. 2012. 09. 033.
[19] ESPOSITO S,DALENO C,SCALA A,et al. Impact of rhinovirus nasopharyngeal viral load and viremia on severity of respiratory infections in children[J]. Eur J Clin Microbiol Infect Dis,2014,33(1):41-48. DOI:10. 1007/s10096-013-1926-5.
[2] JACOBS S E,LAMSON D M,ST GEORGE K,et al. Human rhinoviruses[J]. Clin Microbiol Rev,2013,26(1):135-162. DOI:10. 1128/CMR. 00077-12.
[3]郑首燕.支气管哮喘急性发作患儿鼻病毒检出率及其分子流行病学研究[D].重庆:重庆医科大学,2016.ZHENG S Y. Detection rate of rhinovirus in children with acute asthma attack and its molecular epidemiology[D].Chongqing:Chongqing Medical University,2016.
[4] KANAZAW A J,MASUKO H,YATAGAI Y,et al. Genetic association of the functional CDHR3 genotype with early-onset adult asthma in Japanese populations[J]. Allergol Int,2017,66(4):563-567. DOI:10. 1016/j. alit. 2017. 02. 012.
[5] BOCHKOV Y A,WATTERS K,ASHRAF S,et al. Cadherin-related family member 3,a childhood asthma susceptibility gene product,mediates rhinovirus C binding and replication[J]. Proc Natl Acad Sci USA,2015,112(17):5485-5490. DOI:10. 1073/pnas. 1421178112.
[6] HEIM A,EBNET C,HARSTE G,et al. Rapid and quantitative detection of human adenovirus DNA by real-time PCR[J]. J Med Virol,2003,70(2):228-239. DOI:10. 1002/jmv. 10382.
[7] ALLANDER T,JARTTI T,GUPTA S,et al. Human bocavirus and acute wheezing in children[J]. Clin Infect Dis,2007,44(7):904-910. DOI:10.1086/512196.
[8] COIRAS M T,AGUILAR J C,GARC A M L,et al. Simultaneous detection of fourteen respiratory viruses in clinical specimens by two multiplex reverse transcription nested-PCR assays[J]. J Med Virol,2004,72(3):484-495. DOI:10.1002/jmv. 20008.
[9] COIRAS M T,P REZ-BRE A P,GARC A M L,et al. Simultaneous detection of influenza A,B,and C viruses,respiratory syncytial virus,and adenoviruses in clinical samples by multiplex reverse transcription nested-PCR assay[J]. J Med Virol,2003,69(1):132-144. DOI:10. 1002/jmv. 10255.
[10]肖秋艳.重庆地区人鼻病毒和肠道病毒D68在呼吸道感染住院患儿中的分子流行病学研究[D].重庆:重庆医科大学,2015.XIAO Q Y. Epidemiological analysis of human rhinovirus and enterovirus in hospitalized children with respiratory infections in Chongqing[D]. Chongqing:Chongqing Medical University,2015.
[11] CHANG E H,WILLIS A L,MCCRARY H C,et al. Association between the CDHR3 rs6967330 risk allele and chronic rhinosinusitis[J]. J Allergy Clin Immunol,2017,139(6):1990-1992. e2. DOI:10. 1016/j. jaci. 2016. 10. 027.
[12] ZHAO B,YU X,WANG C,et al. High human bocavirus viral load is associated with disease severity in children under five years of age[J]. PLo S ONE,2013,8(4):e62318. DOI:10. 1371/journal. pone. 0062318.
[13] B NNELYKKE K,SLEIMAN P,NIELSEN K,et al. A genome-wide association study identifies CDHR3 as a susceptibility locus for early childhood asthma with severe exacerbations[J]. Nat Genet,2014,46(1):51-55. DOI:10. 1038/ng. 2830.
[14] BAI J,SMOCK S L,JACKSON G R Jr,et al. Phenotypic responses of differentiated asthmatic human airway epithelial cultures to rhinovirus[J]. PLo S ONE,2015,10(2):e0118286. DOI:10. 1371/journal. pone. 0118286.
[15] STENBERG HAMMAR K, NIESPODZIANA K, VAN HAGE M,et al. Reduced CDHR3 expression in children wheezing with rhinovirus[J]. Pediatr Allergy Immunol,2018,29(2):200-206. DOI:10. 1111/pai. 12858.
[16] HUSBY A,PASANEN A,WAAGE J,et al. CDHR3 gene variation and childhood bronchiolitis[J]. J Allergy Clin Immunol,2017,140(5):1469-1471. e7. DOI:10. 1016/j.jaci. 2017. 06. 044.
[17] LAUINGER I L,BIBLE J M,HALLIGAN E P,et al. Patient characteristics and severity of human rhinovirus infections in children[J]. J Clin Virol,2013,58(1):216-220. DOI:10. 1016/j. jcv. 2013. 06. 042
[18] LINDER J E,KRAFT D C,MOHAMED Y,et al. Human rhinovirus C:age,season,and lower respiratory illness over the past 3 decades[J]. J Allergy Clin Immunol,2013,131(1):69-77. e1-6. DOI:10. 1016/j. jaci. 2012. 09. 033.
[19] ESPOSITO S,DALENO C,SCALA A,et al. Impact of rhinovirus nasopharyngeal viral load and viremia on severity of respiratory infections in children[J]. Eur J Clin Microbiol Infect Dis,2014,33(1):41-48. DOI:10. 1007/s10096-013-1926-5.