Research progress in molecular diagnostic techniques for respiratory tract infections

doi:10.3969/j.issn.1673-8640.2025.10.016

Abstract

Abstract:

In the recent years，the incidence of respiratory infectious diseases has been on the rise. However，traditional diagnostic techniques for respiratory pathogens have limitations，such as long processing time and low determination limit. Molecular diagnostic techniques，on the other hand，offer advantages of rapidity，sensitivity and automation. This review focuses on the role of molecular diagnostic techniques in the etiological diagnosis of respiratory tract infections，and it also looks forward to the future development and direction of molecular diagnostic techniques.

Key words: Respiratory tract infectious disease, Molecular diagnosis, Polymerase chain reaction, Nucleic acid isothermal amplification, Sequencing

CLC Number:

R372

XU Ben, FENG Zhangliang, SONG Xiaojun, JIN Dazhi, CHEN Yu. Research progress in molecular diagnostic techniques for respiratory tract infections[J]. Laboratory Medicine, 2025, 40(10): 1019-1024.

References 33

[1]	国家卫生健康委员会疾病预防控制局. 2021年全国法定传染病疫情概况[J]. 中国病毒病杂志, 2022, 12(3):236.
[2]	郑泽扬, 王众楷, 王连珂, 等. 2004―2018年中国主要呼吸道传染病的流行趋势及特征[J]. 中华疾病控制杂志, 2022, 26(6):624-630.
[3]	郭彦彤. 基于荧光定量PCR的呼吸道病原体核酸快检技术研究[D]. 广州: 南方医科大学, 2023.
[4]	宋晓冬, 曹国君, 朱旻, 等. 两种分子POCT产品在流感病毒核酸快速诊断中的应用评价[J]. 国际检验医学杂志, 2024, 45(2):146-149.
[5]	应春, 沈彩燕, 詹建华, 等. HMGB1对呼吸道合胞病毒毛细支气管炎患儿单核细胞免疫功能的影响[J]. 中国病理生理杂志, 2022, 38(8):1493-1498.
[6]	曾令文, 蔡林再, 陈贤胜, 等. miR-183-5p通过线粒体凋亡途径影响肺炎支原体诱导的RAW246.7细胞凋亡的机制研究[J]. 中国免疫学杂志, 2023, 39(6):1248-1251.
[7]	李虎虎, 王宝强, 王原媛, 等. 多重荧光PCR法和免疫荧光法诊断常见急性呼吸道病毒感染的一致性研究[J]. 标记免疫分析与临床, 2022, 29(9):1528-1531.
[8]	WANG B H, LIN J C, HUANG H N, et al. Comprehensive evaluation of an one pot quadruplex RT-qPCR assay for the simultaneously subtyping influenza A virus into H1,H3,N1 and N2[J]. J Clin Virol Plus, 2022, 2(4):110-119.
[9]	LI Q, WU D, LI R, et al. Valproic acid protects neurons and promotes neuronal regeneration after brachial plexus avulsion[J]. Neural Regen Res, 2013, 8(30):2838-2848. DOI PMID
[10]	年悬悬, 周辉, 李军英, 等. 乙型流感病毒荧光定量RT-PCR检测方法的建立及验证[J]. 中国生物制品学杂志, 2022, 35(9):1090-1095.
[11]	NGUYEN T T, KWON H J, KIM I H, et al. Multiplex nested RT-PCR for detecting avian influenza virus,infectious bronchitis virus and Newcastle disease virus[J]. J Virol Methods, 2013, 188(1):41-46. DOI URL
[12]	ARVIA R, CORCIOLI F, SIMI L, et al. Monitoring the susceptibility to oseltamivir of Influenza A(H1N1)2009 virus by nested-PCR and pyrosequencing during the pandemic and in the season 2010-2011[J]. J Virol Methods, 2012, 184(1):113-116. DOI URL
[13]	WANG M, YANG J, GAI Z, et al. Comparison between digital PCR and real-time PCR in detection of Salmonella typhimurium in milk[J]. Int J Food Microbiol, 2018, 266:251-256. DOI URL
[14]	COUDRAY-MEUNIER C, FRAISSE A, MARTIN-LATIL S, et al. A comparative study of digital RT-PCR and RT-qPCR for quantification of Hepatitis A virus and Norovirus in lettuce and water samples[J]. Int J Food Microbiol, 2015, 201:17-26. DOI URL
[15]	池细俤, 高世华, 张忠源, 等. 环介导等温扩增技术检测呼吸道病毒方法的建立及应用[J]. 中国实验诊断学, 2021, 25(9):1268-1272.
[16]	罗思思, 谢芝勋, 谢丽基, 等. H7亚型和N9亚型禽流感病毒RT-LAMP可视化检测方法的建立[J]. 畜牧兽医学报, 2015, 46(7):1176-1183.
[17]	PARK B H, OH S J, JUNG J H, et al. An integrated rotary microfluidic system with DNA extraction,loop-mediated isothermal amplification,and lateral flow strip based detection for point-of-care pathogen diagnostics[J]. Biosens Bioelectron, 2017, 91:334-340. DOI URL
[18]	AUGUSTINE R, HASAN A, DAS S, et al. Loop-mediated isothermal amplification(LAMP):a rapid,sensitive,specific,and cost-effective point-of-care test for coronaviruses in the context of COVID-19 pandemic[J]. Biology(Basel), 2020, 9(8):182.
[19]	孙弦, 刘睿, 高虎, 等. NASBA与RT-qPCR技术检测HIV阴性马尔尼菲篮状菌病患者病灶组织真菌载量的效果及相关性[J]. 广西医学, 2023, 45(11):1361-1364.
[20]	蒲姝旸, 张瑾逸, 暴晓阳, 等. 梨火疫病菌的RPA快速检测技术的研发[J]. 植物保护, 2024, 50(4):242-249.
[21]	杨艳兵, 胡海英, 吴茱萸, 等. 基因芯片技术在成人下呼吸道感染病原学检测中的临床应用及评价[J]. 广东医学, 2019, 40(21):3025-3029.
[22]	崔鹤馨. 3种呼吸道病毒基因芯片检测方法的建立与研究[D]. 长春: 吉林大学, 2013.
[23]	陈榕方, 吴雁. 微流控芯片技术在临床医学检测中的应用进展[J]. 医疗装备, 2022, 35(20):191-194.
[24]	任琴, 张星艳, 杜寿文, 等. 基于微流控芯片技术的五种呼吸道病毒核酸快速荧光qPCR检测方法的建立[J]. 新发传染病电子杂志, 2020, 5(2):95-99.
[25]	胡付品, 郭燕, 朱德妹, 等. 2021年CHINET中国细菌耐药监测[J]. 中国感染与化疗杂志, 2022, 22(5):521-530. DOI
[26]	韩心远, 高小娟. 宏基因组测序技术在感染性病原体检测中的应用进展[J]. 海南医学, 2022, 33(14):1875-1878.
[27]	林彦锋. 宏基因组测序技术在急性呼吸道感染病原识别中的应用研究[D]. 北京: 中国人民解放军军事科学院, 2020.
[28]	MITCHELL S L, SIMNER P J. Next-generation sequencing in clinical microbiology:are we there yet?[J]. Clin Lab Med, 2019, 39(3):405-418. DOI URL
[29]	刘滕颖子, 刘南希, 张晓敏, 等. 靶向测序综合分析诊断结核分枝杆菌合并多病原体感染肺炎1例[J]. 新发传染病电子杂志, 2024, 9(3):73-77. DOI
[30]	廖毓香, 朱水泉, 伍桂雄, 等. 靶向高通量测序技术(tNGS)在下呼吸道感染病原体检测中的诊断价值[J]. 系统医学, 2024, 9(6):81-83.
[31]	叶福强, 李鹏, 韩一芳, 等. 基于纳米孔测序技术的呼吸道病原体快速确认[J]. 分子诊断与治疗杂志, 2020, 12(2):127-132.
[32]	CONG L, RAN F A, COX D, et al. Multiplex genome engineering using CRISPR/Cas systems[J]. Science, 2013, 339(6121):819-823. DOI PMID
[33]	ZHOU H, TSOU J H, CHINTHALAPALLY M, et al. Detection and differentiation of SARS-CoV-2,influenza,and respiratory syncytial viruses by CRISPR[J]. Diagnostics(Basel), 2021, 11(5):823.