[1]唐月明,伊 洁.数字聚合酶链反应(dPCR)技术在病原体基因检测应用中的研究进展[J].现代检验医学杂志,2021,36(05):174-179.[doi:10.3969/j.issn.1671-7414.2021.05.038]
 TANG Yue-ming,YI Jie.Recent Advances in Research on Digital Polymerase ChainReaction(dPCR) in Pathogen Gene Detection[J].Journal of Modern Laboratory Medicine,2021,36(05):174-179.[doi:10.3969/j.issn.1671-7414.2021.05.038]
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数字聚合酶链反应(dPCR)技术在病原体基因检测应用中的研究进展()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第36卷
期数:
2021年05期
页码:
174-179
栏目:
综 述
出版日期:
2021-10-14

文章信息/Info

Title:
Recent Advances in Research on Digital Polymerase ChainReaction(dPCR) in Pathogen Gene Detection
文章编号:
1671-7414(2021)05-174-06
作者:
唐月明伊 洁
(中国医学科学院北京协和医院检验科,北京 100730)
Author(s):
TANG Yue-ming YI Jie
(Department of Clinical Laboratory, Peking Union Medical College Hospital, ChineseAcademy of Medical Science, Beijing 100730, China)
关键词:
数字聚合酶链反应病原体绝对定量
分类号:
Q503
DOI:
10.3969/j.issn.1671-7414.2021.05.038
文献标志码:
A
摘要:
实时荧光定量聚合酶链反应(qPCR)的准确度和精密度容易受到很多因素的影响,数字聚合酶链反应(dPCR)作为一种新的分子定量技术,以其更高的敏感度、精密度、独立于标准曲线及更高的抗干扰能力等优点,近年来在病原体基因检测中逐渐兴起。该文将dPCR技术的特点、优势、不足及在病原体检测中的应用进行综述。
Abstract:
The accuracy and precision of real-time quantitative polymerase chain reaction (qPCR) are susceptible to many factors.Digital polymerase chain reaction (dPCR), as a new molecular quantitative technique, has emerged in pathogen gene detectionin recent years with advantages of higher sensitivity, precision, independence from standard curves and higher anti-interferenceability. This article reviews the characteristics, progress and applications of dPCR in pathogen gene detection of pathogen, and itsprospect in the future .

参考文献/References:

[1] KUYPERS J, JEROME K R. Applications of digitalPCR for clinical microbiology[J]. Journal of ClinicalMicrobiology, 2017, 55(6): 1621-1628.
[2] VOGELSTEIN B, KINZLER K W. Digital PCR[J].Proceedings of the National Academy of Sciences ofthe United States of America, 1999, 96(16): 9236-9241.
[3] VYNCK M, TRYPSTEEN W, THAS O, et al.The future of digital polymerase chain reaction invirology[J]. Molecular Diagnosis & Therapy, 2016,20(5): 437-447.
[4] CAO Lei, CUI Xingye, HU Jie, et al. Advances indigital polymerase chain reaction (dPCR) and itsemerging biomedical applications [J]. Biosensors &Bioelectronics, 2017, 90: 459-474.
[5] CAO Guojun, TAN Chen, ZHANG Yuyan,et al. Digitaldroplet polymerase chain reaction analysis of commonviruses in the aqueous humour of patients with Posner-Schlossman syndrome in Chinese population[J].Clinical & Experimental Ophthalmology, 2019, 47(4):513-520.
[6] ZHANG Kuo, LIN Guigao, LI Jinming. Quantitativenucleic acid amplification by digital PCR for clinicalviral diagnostics[J]. Clinical Chemistry and LaboratoryMedicine, 2016, 54(9): 1427-1433.
[7] SIDSTEDT M, R?DSTR?M P, HEDMAN J. PCRinhibition in qPCR, dPCR and MPS-mechanisms andsolutions[J]. Analytical and Bioanalytical Chemistry,2020, 412(9): 2009-2023.
[8] TANG Li, SUN Yilun, BUELOW D, et al. Quantitativeassessment of commutability for clinical viral loadtesting using a digital PCR-based reference standard[J].Journal of Clinical Microbiology, 2016, 54(6): 1616-1623.
[9] QUAN P L ,SAUZADE M, BROUZES E. dPCR:A technology review [J]. Sensors (Basel), 2018,18(4):1271.
[10] NIXON G, GARSON J A, GRANT P, et al. Comparativestudy of sensitivity, linearity, and resistance to inhibitionof digital and nondigital polymerase chain reactionand loop mediated isothermal amplification assays forquantification of human cytomegalovirus[J]. AnalyticalChemistry, 2014, 86(9): 4387-4394.
[11] KOJABAD A A, FARZANEHPOUR M, GOUVARCHINGALEH H E, et al. Droplet digital PCR of viral DNA/RNA, current progress, challenges, and future perspectives[J]. Journal of Medical Virology, 2021. DOI:10.1002/jmv.26846.
[12] USHIO R, YAMAMOTO M, NAKASHIMA K,et al. Digital PCR assay detection of circulatingMycobacterium tuberculosis DNA in pulmonarytuberculosis patient plasma[J]. Tuberculosis, 2016, 99 :47-53.
[13] JEANNOT E, BECETTE V, CAMPITELLI M, et al.Circulating human papillomavirus DNA detected usingdroplet digital PCR in the serum of patients diagnosedwith early stage human papillomavirus-associatedinvasive carcinoma[J]. The Journal of Pathology.Clinical Research, 2016, 2(4): 201-209.
[14] PERSAUD D, GAY H, ZIEMNIAK C, et al. Absenceof detectable HIV-1 viremia after treatment cessationin an infant[J]. The New England Journal of Medicine,2013, 369(19): 1828-1835.
[15] GUPTA R K, PEPPA D, HILL A L, et al. Evidencefor HIV-1 cure after CCR5Δ32/Δ32 allogeneichaemopoietic stem-cell transplantation 30 months postanalytical treatment interruption: a case report[J]. TheLancet HIV, 2020, 7(5): e340-e347.
[16] WAGGONER J, HO D Y, LIBIRAN P, et al. Clinicalsignificance of low cytomegalovirus DNA levels inhuman plasma[J]. Journal of Clinical Microbiology,2012, 50(7): 2378-2383.
[17] LIMAYE A P, BABU T M, BOECKH M. Progress andchallenges in the prevention, diagnosis and managementof cytomegalovirus infection in transplantation[J].Clinical Microbiology Reviews, 2020, 34(1): e00043-19.
[18] POMARI E, PIUBELLI C, PERANDIN F, et al. DigitalPCR: a new technology for diagnosis of parasiticinfections[J]. Clinical Microbiology and Infection,2019, 25(12): 1510-1516.
[19] SEDLAK R H, HILL J A, NGUYEN T, et al. Detectionof human herpesvirus 6B (HHV-6B) reactivation inhematopoietic cell transplant recipients with inheritedchromosomally integrated HHV-6A by droplet digitalPCR[J]. Journal of Clinical Microbiology, 2016, 54(5):1223-1227.
[20] LIMOTHAI U, CHUAYPEN N, POOVORAWANK, et al. Reverse transcriptase droplet digital PCR vsreverse transcriptase quantitative real-time PCR forserum HBV RNA quantification[J]. Journal of MedicalVirology, 2020,92(12): 3365-3372.
[21] DICKSON R P, SCHULTZ M J, VAN DER POLLT, et al. Lung microbiota predict clinical outcomesin critically ill patients[J]. American Journal ofRespiratory and Critical Care Medicine, 2020, 201(5):555-563.
[22] LIU Chong, SHI Qingxin, PENG Mingfei, et al.Evaluation of droplet digital PCR for quantificationof SARS-CoV-2 virus in discharged COVID-19patients[J]. Aging, 2020, 12(21): 20997-21003.
[23] YU Fengting, YAN Liting, WANG Nan, et al.Quantitative detection and viral load analysis of SARSCoV-2 in infected patients[J]. Clinical InfectiousDiseases, 2020, 71(15): 793-798.
[24] SUO Tao, LIU Xinjin, FENG Jiangpeng, et al. ddPCR:a more accurate tool for SARS-CoV-2 detection inlow viral load specimens[J]. Emerging Microbes &Infections, 2020, 9(1): 1259-1268.
[25] PEKIN D, SKHIRI Y, BARET J C, et al. Quantitativeand sensitive detection of rare mutations using dropletbasedmicrofluidics[J]. Lab on a Chip, 2011, 11(13):2156-2166.
[26] BATEMAN A C, GRENINGER A L, ATIENZA E E, etal. Quantification of BK virus standards by quantitativereal-time PCR and droplet digital PCR is confoundedby multiple virus populations in the WHO BKVinternational standard[J]. Clinical Chemistry, 2017,63(3): 761-769.
[27] GRENINGER A L, BATEMAN A C, ATIENZA E E, etal. Copy number heterogeneity of JC virus standards[J].Journal of Clinical Microbiology, 2017, 55(3): 824-831.
[28] LUO Jun, LI Junhua, YANG Hang, et al. Accuratedetection of methicillin-resistant Staphylococcus aureusin mixtures by use of single-bacterium duplex dropletdigital PCR[J]. Journal of Clinical Microbiology, 2017,55(10): 2946-2955.
[29] ABRAM T J, CHERUKURY H, OU C Y, et al. Rapidbacterial detection and antibiotic susceptibility testingin whole blood using one-step, high throughput blooddigital PCR[J]. Lab on a Chip, 2020, 20(3): 477-489.
[30] HAYNES R J, KLINE M C, TOMAN B, et al. Standardreference material 2 366 for measurement of humancytomegalovirus DNA[J]. Journal of MolecularDiagnostics, 2013, 15(2): 177-185.
[31] PAV?I? J, DEVONSHIRE A, BLEJEC A, et al.Inter-laboratory assessment of different digital PCRplatforms for quantification of human cytomegalovirusDNA[J]. Analytical and Bioanalytical Chemistry, 2017,409(10): 2601-2614.
[32] TALARICO S, SAFAEIAN M, GONZALEZ P, et al.Quantitative detection and genotyping of Helicobacterpylori from stool using droplet digital PCR revealsvariation in bacterial loads that correlates with cagAvirulence gene carriage[J]. Helicobacter, 2016, 21(4):325-333.
[33] WU Xulong, LIN Hua, CHEN Shijie, et al.Development and application of a reverse transcriptasedroplet digital PCR (RT-ddPCR) for sensitive and rapiddetection of Japanese encephalitis virus[J]. Journal ofVirological Methods, 2017, 248:166-171.
[34] ROSALES-CHILAMA M, DIAZ-MORENO N,PRIETO M D, et al. Comparative assessment of DNAtargets and amplification methods for leishmania(viannia) detection in human samples[J]. The AmericanJournal of Tropical Medicine and Hygiene, 2020,102(6): 1323-1327.
[35] DING Yun, CHOO J, DE MELLO A J. From singlemoleculedetection to next-generation sequencing:microfluidic droplets for high-throughput nucleic acidanalysis[J]. Microfluidics and Nanofluidics, 2017,21(3): 58.
[36] EASTBURN D J, HUANG Y, PELLEGRINO M,et al. Microfluidic droplet enrichment for targetedsequencing[J]. Nucleic Acids Research, 2015, 43(13):e86.
[37] YU Ziqing, L? Weiyuan,YU Mengchao, et al. SelfpartitioningSlipChip for slip-induced droplet formationand human papillomavirus viral load quantificationwith digital LAMP[J]. Biosensors & Bioelectronics,2020, 155: 112107.
[38] GORGANNEZHAD L, STRATTON H, NGUYEN N T.Microfluidic-Based nucleic acid amplification systemsin microbiology[J]. Micromachines, 2019, 10(6): 408.
[39] HAYDEN R T, GU Z, SAM S S, et al. Comparativeperformance of reagents and platforms for quantitationof cytomegalovirus DNA by digital PCR[J]. Journal ofClinical Microbiology, 2016, 54(10): 2602-2608.

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备注/Memo

备注/Memo:
基金项目: “十三五”艾滋病和病毒性肝炎等重大传染病防治科技重大专项“课题”急性呼吸道传染病病原谱和流行规律及重要病原变异变迁特征研究(2017ZX10103004003)。
作者简介:唐月明(1992-),女,硕士,技师,主要从事病原微生物检测,E-mail:mingming-10100601@163.com。
更新日期/Last Update: 1900-01-01