Establishment of fluorescent quantitative polymerase chain reaction detection method for human parvovirus B19

XU Liangyun; ZHANG Qigang; FAN Guanglai; ZHAO Jianhua

doi:10.7619/jcmp.20210246

Journal of Clinical Medicine in Practice > 2021 > 25(7): 11-14, 20. > DOI: 10.7619/jcmp.20210246

XU Liangyun, ZHANG Qigang, FAN Guanglai, ZHAO Jianhua. Establishment of fluorescent quantitative polymerase chain reaction detection method for human parvovirus B19[J]. Journal of Clinical Medicine in Practice, 2021, 25(7): 11-14, 20. DOI: 10.7619/jcmp.20210246

Citation:

PDF (1997 KB)

Establishment of fluorescent quantitative polymerase chain reaction detection method for human parvovirus B19

1.
Tumor Hospital Affiliated to Fourth Clinical Medical College of Nanjing Medical University, Nanjing, Jiangsu, 210009
2.
Gene Testing Technology Application Demonstration Center, Huai'an City Maternal and Child Health Hospital of Jiangsu Province, Huai'an, Jiangsu, 223002

More Information

Received Date: January 12, 2021
Available Online: April 20, 2021
Published Date: July 27, 2021

Graphical Abstract

Abstract

Abstract

Objective To establish a fluorescent quantitative polymerase chain reaction detection method (PCR) method for simultaneous detection of human parvovirus B19(HPV B19).
Methods The online tool MAFFT was used to compare and analyze the sequence NS1 gene conserved region of three genotypes HPV B19, and specific primers and probes were designed for conserved region sequences. The fluorescent quantitative PCR method for detecting HPV B19 was established, and the sensitivity and specificity of the method were evaluated.
Results A standard curve drawn with a plasmid DNA template at a concentration of 1×10⁷ to 1×10² copies/μL and 20 copies/μL, the reaction efficiency E was 0.946 and R² was 0.996 6; the detection limit of this method was 10 copies/μL and the lower limit of quantification was 20 copies/μL. Herpes simplex virus (HSV) type 2, cytomegalovirus (CMV), group B streptococcus (GBS), hepatitis B virus (HBV), Epstein-Barr virus (EBV) positive DNA virus test results were negative, and the reference plasmid and HPV B19 positive samples were amplified.
Conclusion The established fluorescent quantitative PCR detection method can be applied for the screening and diagnosis of HPV B19 infection as high sensitivity and specificity.
- human parvovirus B19,
- fluorescent quantitative polymerase chain reaction,
- standard curve,
- sequence

FullText(HTML)

References (15)

References

[1]	PAVER W K, CAUL E O, CLARKE S K. Letter: Parvovirus-like particles in human sera[J]. Lancet, 1975, 1(7900): 232-232. http://europepmc.org/abstract/med/46024
[2]	WALDMAN M, KOPP J B. Parvovirus B19 and the kidney[J]. Clin J Am Soc Nephrol, 2007, 2(Suppl 1): S47-S56. http://www.ncbi.nlm.nih.gov/pubmed/17699510
[3]	KAHN J S, KESEBIR D, COTMORE S F, et al. Seroepidemiology of human bocavirus defined using recombinant virus-like particles[J]. J Infect Dis, 2008, 198(1): 41-50. doi: 10.1086/588674
[4]	DE JONG E P, WALTHER F J, KROES A C M, et al. Parvovirus B19 infection in pregnancy: new insights and management[J]. Prenat Diagn, 2011, 31(5): 419-425. doi: 10.1002/pd.2714
[5]	ROGO L D, MOKHTARI-AZAD T, KABIR M H, et al. Human parvovirus B19: a review[J]. Acta Virol, 2014, 58(3): 199-213. doi: 10.4149/av_2014_03_199
[6]	罗青清, 骆名恋, 邹丽. "妊娠期微小病毒B19感染的临床实践指南"解读[J]. 中国实用妇科与产科杂志, 2016, 32(6): 502-504. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGSF201606004.htm
[7]	VALEUR-JENSEN A K, PEDERSEN C B, WESTERGAARD T, et al. Risk factors for parvovirus B19 infection in pregnancy[J]. JAMA, 1999, 281(12): 1099-1105. doi: 10.1001/jama.281.12.1099
[8]	HARGER J H, ADLER S P, KOCH W C, et al. Prospective evaluation of 618 pregnant women exposed to parvovirus B19: risks and symptoms[J]. Obstet Gynecol, 1998, 91(3): 413-420. doi: 10.1016/S0029-7844(97)00701-1
[9]	ZHOU Y, BIAN G H, ZHOU Q X, et al. Detection of cytomegalovirus, human parvovirus B19, and Herpes simplex virus-1/2 in women with first-trimester spontaneous abortions[J]. J Med Virol, 2015, 87(10): 1749-1753. doi: 10.1002/jmv.24218
[10]	杨忠心, 何惠君, 代方, 等. 无偿献血员血清人微小病毒B19感染情况调查[J]. 华中科技大学学报: 医学版, 2003, 32(3): 344-345. doi: 10.3870/j.issn.1672-0741.2003.03.035
[11]	JIA J T, MA Y Y, ZHAO X, et al. Prevalence of human parvovirus B19 in Chinese plasma pools for manufacturing plasma derivatives[J]. Virol J, 2015, 12: 162-162. doi: 10.1186/s12985-015-0396-z
[12]	BREDL S, PLENTZ A, WENZEL J J, et al. False-negative serology in patients with acute parvovirus B19 infection[J]. J Clin Virol, 2011, 51(2): 115-120. doi: 10.1016/j.jcv.2011.03.012
[13]	JIA J T, ZHONG Y D, GUO Y, et al. Simultaneous detection and differentiation of human parvovirus B19 and human parvovirus 4 by an internally controlled multiplex quantitative real-time PCR[J]. Mol Cell Probes, 2017, 36: 50-57. doi: 10.1016/j.mcp.2017.08.005
[14]	ALVES A D R, CUBEL GARCIA R C N, CRUZ O G, et al. Quantitative real-time PCR for differential diagnostics of parvovirus B19 infection in acute liver failure patients[J]. Expert Rev Mol Diagn, 2019, 19(3): 259-266. doi: 10.1080/14737159.2019.1582333
[15]	VON KIETZELL K, POZZUTO T, HEILBRONN R, et al. Antibody-mediated enhancement of parvovirus B19 uptake into endothelial cells mediated by a receptor for complement factor C1q[J]. J Virol, 2014, 88(14): 8102-8115. doi: 10.1128/JVI.00649-14

[1]	GAO Nairong, GUO Jingrui. Research on budesonide aerosol inhalation in treatment of children with asthma[J]. Journal of Clinical Medicine in Practice, 2020, 24(14): 63-66. DOI: 10.7619/jcmp.202014017
[2]	FAN Wenjuan, WU Shengmei. Effect of evidence-based nursing on the efficacy and family satisfaction of asthma children with atomization inhalation[J]. Journal of Clinical Medicine in Practice, 2016, (14): 140-143. DOI: 10.7619/jcmp.201614044
[3]	ZHONG Yueying. Application of whole-course nursing intervention in atomization inhalation treatment for children with bronchial asthma[J]. Journal of Clinical Medicine in Practice, 2016, (6): 123-125. DOI: 10.7619/jcmp.201606039
[4]	XIE Yumei. Application of health education in patients with bronchial asthma treated by atomization inhalation[J]. Journal of Clinical Medicine in Practice, 2016, (6): 120-122. DOI: 10.7619/jcmp.201606038
[5]	FU Hong, LI Wei, LIU Jing. Effect of cluster nursing in children with pneumonia treated by atomization inhalation[J]. Journal of Clinical Medicine in Practice, 2016, (6): 117-119. DOI: 10.7619/jcmp.201606037
[6]	ZHU Yingxia. Nursing measures of oxygen aerosol inhalation in the treatment of children with moderate or severe asthma[J]. Journal of Clinical Medicine in Practice, 2016, (2): 115-117,121. DOI: 10.7619/jcmp.201602036
[7]	LI Yi, ZHAI Hong, XIAO Xueqing. Effects of budesonide suspension by oxygen atomization inhalation in treatment of child with acute paroxysm of asthma[J]. Journal of Clinical Medicine in Practice, 2015, (Z1): 64-65.
[8]	LIU Dongmei, ZHU Ying. Control study on mechanical sputum aspiration combined with atomization inhalation in patients with respiratory failure[J]. Journal of Clinical Medicine in Practice, 2015, (22): 60-63. DOI: 10.7619/jcmp.201522020
[9]	DONG Lingzhi, REN Zhiyan, DONG Shuling. Efficacy and nursing of aerosol inhalation therapy of budesonide suspension combined with compound ipratropium bromide in the treatment of children with asthma[J]. Journal of Clinical Medicine in Practice, 2015, (8): 97-100. DOI: 10.7619/jcmp.201508031
[10]	TANG Fujie. Effect observation of different atomizing inhalation therapy on treatment of acute laryngitis children with laryngeal obstruction[J]. Journal of Clinical Medicine in Practice, 2014, (15): 66-69. DOI: 10.7619/jcmp.201415020

Cited By

Cited by

Periodical cited type(5)

1.	刘培彩，陈倩，苏杨. 穴位贴敷联合电子无烟艾灸对慢性心力衰竭患者心脏功能及日常生活能力的影响. 中国医药指南. 2025(07): 156-158 .
2.	黄雨欣，杨帆，陈奇祺，廖佳颖，李潇杰，黎祖鸣，陈慧聪. 实脾散临床应用研究进展. 中国民族民间医药. 2024(03): 76-80 .
3.	程虹，王长江，邹新蓉，薛雪，王小琴. 王小琴分期论治糖尿病肾病经验管窥. 中国医药导报. 2024(04): 143-147 .
4.	张运娇，杨继，马腾，田昕彤，穆丽婷，赵英强. 经方治疗心力衰竭的研究现状. 中西医结合心脑血管病杂志. 2023(08): 1444-1450 .
5.	史振红，周红妮，崔东. 实脾饮合肾气丸治疗慢性肾功能衰竭. 中医学报. 2022(09): 1977-1981 .

Other cited types(5)

Get Citation

PDF

XML

Article views (523) PDF downloads (41) Cited by(10)

Turn off MathJax

Article Contents

Abstract

References

Establishment of fluorescent quantitative polymerase chain reaction detection method for human parvovirus B19