色谱 ›› 2020, Vol. 38 ›› Issue (9): 999-1012.DOI: 10.3724/SP.J.1123.2020.02029

• 专论与综述 • 上一篇    下一篇

毛细管电泳技术在疾病预防控制领域的应用、发展与挑战

林长缨, 丁晓静()   

  1. 北京市疾病预防控制中心, 食物中毒溯源技术北京市重点实验室, 北京市预防医学研究中心, 北京 100013
  • 收稿日期:2020-02-29 出版日期:2020-09-08 发布日期:2020-12-11
  • 通讯作者: 丁晓静
  • 作者简介:丁晓静.Tel:(010)64407087, E-mail:dxj666@aliyun.com
  • 基金资助:
    国家重点研发计划(2016YFF0201106)

Application, development, and challenges of capillary electrophoresis in disease prevention and control

LIN Changying, DING Xiaojing()   

  1. Beijing Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning, Beijing Center for Disease Control and Prevention, Beijing Research Center for Preventive Medicine, Beijing 100013, China
  • Received:2020-02-29 Online:2020-09-08 Published:2020-12-11
  • Contact: DING Xiaojing
  • Supported by:
    National Key R&D Program of China(2016YFF0201106)

摘要:

自1989年出现商品化的仪器以来,毛细管电泳(CE)技术在多个应用领域都取得了长足的进步与发展,重复性和准确性方面也有很大提升。能力验证样品分析的满意结果也显示了CE具备法规要求的准确定量能力。在疾病预防控制领域(简称"疾控")CE也展现出很多独具特色的应用,成为不可或缺的技术之一。在聚合酶链式反应产物分析、核酸序列测定、DNA变异和分型分析、食源性致病微生物分析及疫苗分析等工作中CE发挥了重要作用。应对突发疫情或公共卫生事件如食物中毒时,除了通过非靶标分析尽快锁定目标物外,还需要对大量样品做出快速而准确的分析,高通量和高灵敏的CE就十分适合解决这一问题。在公共卫生理化检验以确保食品、保健食品、特殊医学用途食品、化妆品和消毒产品等的安全中,CE也发挥了不可或缺的作用。作为一种使用较少危险化学品的环境友好方法,在需要按照标准或规范进行的疾控实验室常规检测中,CE仍受制于标准方法的缺失,未能发挥其应有的作用。但简单、快速、经济、耐用、高效的CE分离一旦与高灵敏通用检测器联用,必将更加从容地应对疾控领域中的各种挑战,发挥更大的作用。本文综述了2010~2019年CE在疾控领域的应用,分析了CE在疾控领域发展的机遇和挑战,对CE在疾控领域的发展前景进行了展望。

关键词: 毛细管电泳, 核酸, 病原微生物, 疫苗, 疾病预防控制, 能力验证, 食物中毒, 应急分析

Abstract:

Since the advent of commercial instruments in 1989, capillary electrophoresis (CE) has advanced considerably, with improvement in reproducibility and accuracy in many application fields. CE is predominantly used in research on disease prevention and control, and hygienic chemical inspection. The applications of CE range from assessment of inorganic anions and cations in drinking water to that of biological macromolecules, such as nucleic acids, in pathogenic microorganisms. Since the analytical capacity of CE ranges from inorganic ions to cell, it has become an indispensable technique in this field, particularly in public health emergency and epidemic management. Universal non-targeted analyses to detect possible pathogens, and the capability of rapid and accurate testing of large numbers of specimens are required. In the analyses of polymerase chain reaction (PCR) products, nucleic acid sequencing, mutation detection and genotyping, food-borne disease pathogens, and vaccine analyses, CE methods characterized by high through-put and sensitivity are necessary. In the public health sector, CE is essential in the analyses of food (including emergency analyses for food poisoning), cosmetics, and disinfectants. Satisfactory results of the FAPAS (Food Analysis Performance Assessment Scheme) and domestic proficiency tests indicated the accuracy of CE in quantitative analyses. Application of CE in disease prevention and control is challenged by a number of new molecular biological methods, as optimizing CE methods may not be feasible, and results are difficult to interpret. CE methods, including transformation of peaks to identification of pathogens, can be an arduous task. Thus, end-users prefer using commercialized CE systems and reagents in their routine work. Alternatively, CE methods for analysis of small molecules in product analyses, such as food safety, cosmetics and disinfectant testing, is commonly performed. A plethora of studies published within the decade, indicate that CE is still an active research area in hygienic chemical inspection. To a large extent, CE has not been used for routine analysis in the centers for disease control and prevention, accredited laboratories in China, nor regulatory agencies worldwide. This may be due to the lack of practical protocols for the standards, and the misconceptions regarding the ease of use of CE, which could have hindered its widespread application. Although CE is an environmental friendly technique with minimal usage of toxic chemicals, few standard methods of CE exist in agriculture, environmental protection, food, beverage, chemical, and pharmaceutical industries in the United States, Britain, Europe, Japan, India, Brazil, Russia, and China. Since 2002, CE was used in our laboratory to analyze a large variety of samples. We found that once the CE method has been fully verified and described in detail, it was easily standardized. It is not necessary to screen the equivalent chromatographic column, or to use a specific liquid chromatographic (LC) column. This can effectively circumvent the challenge of shifting peak orders caused by different LC column selectivity. Once combined with general, high sensitivity detectors, CE can be used in the detection of bacteria or viruses in food safety, and play a greater role in the field of disease prevention and control. In the present review, applications of CE in nucleic acid detection for viruses and bacteria, analysis of vaccines, routine testing on food, dietary supplements, medical foods, cosmetics and disinfectants, proficiency tests, and emergency analyses of food poisoning were summarized. The applications and challenges of CE in the field of disease control and prevention were analyzed, and development of this technique was prospected.

Key words: capillary electrophoresis (CE), nucleic acid, pathogen microbes, vaccine, disease control and prevention, proficiency test, food poisoning, emergency analysis