基于富集技术的医院大面积物表病原采样处理方法

Research on etiological sampling for large-scale object surfaces in hospitals based on enrichment technology

    摘要
  • 摘要:
    目的 对比传统拭子采样法在医院大面积物表传染病病原监测中的效果，验证基于预湿抗静电布与改良聚二乙醇(PEG)沉淀富集技术的采样处理方法，以提升传染病早期预警能力及优化医院环境监测方案。
    方法 2024年5月-2025年3月，在深圳市龙岗人民医院、龙岗区第二人民医院和龙岗区妇幼保健院3家公立医院开展8次现场监测，采用荧光定量聚合酶链反应(PCR)同时检测23种呼吸道病原(18种病毒、5种致病菌)及6种胃肠道病原，比较实际检出率、病原谱及循环阈值(Ct)；实验室评价以不同载量H1N1流感病毒加标亚克力板为模型，对比两种方法的最低检测限、灵敏度及重复性。
    结果 两种方法最低检测限均为6.0×104 copies/ml，但预湿布法核酸检测阳性率(100.00%，3/3)高于拭子法(33.33，1/3)；低病毒载量(6.0×105 copies/ml)时，预湿布法平均病毒核酸浓度(X-Ct=36.59)更高，重复性更优(CV=0.99%，＜3.14%)。现场监测中，预湿布法病原总体检出率(42.84%~64.27%)高于拭子法(10.71%~21.43%)，检出病原更丰富，Ct值更低(P＜0.05)。
    结论 预湿布采样富集法整合抗静电布采样与PEG富集技术，在大面积物表病原监测中灵敏度和稳定性更高，可提高检出率。

     

    Abstract:
    OBJECTIVE To compare the effect of traditional swab sampling method on etiological surveillance of infectious diseases for large-scale object surfaces in hospitals and validate the samples processing method based on pre-wet anti-static fabric and modified polyethylene glycol (PEG) precipitation enrichment technology so as to improve the capability of early warning of infectious diseases and optimize the environmental surveillance program in the hospitals.
    METHODS The on-site surveillance was carried out for 8 times in 3 public hospitals (Shenzhen Longgang People′s Hospital, the Second People′s Hospital of Longgang and Longgang Maternal and Child Health Hospital) from May 2024 to Mar. 2025. Totally 23 types of respiratory tract pathogens (18 types of viruses, 5 type of pathogenic bacteria) and 6 types of gastrointestinal tract pathogens were simultaneously detected by means of fluorescent quantitative polymerase chain reaction (PCR); the actual isolation rates, etiological spectrum and cycle threshold (Ct) value were compared. The acrylic plate added with standards of different loads of H1NI influenza viruses was used as model for laboratory evaluation. The minimum detection limit, sensitivity and repeatability were observed and compared between the methods.
    RESULTS The minimum detection limit of both methods was 6.0×104 copies/ml, however, the positive rate of nucleic acid testing of the pre-wet fabric method was 100.00% (3/3), higher than 33.33% (1/3) of the swab method; when the low viral load was 6.0×105 copies/ml, the average concentration of viral nucleic acid of the pre-wet fabric method (X-Ct=36.59) was higher, with the repeatability (CV=0.99%, < 3.14%) better. The results of the on-site surveillances showed that the total isolation rates of pathogens of the pre-wet fabric method ranged between 42.84% and 64.27%, higher than between 10.71% and 21.43% of the swab method, with the isolated pathogens more abundant, the Ct value lower(P < 0.05).
    CONCLUSION The pre-wet fabric sampling enrichment method integrated with anti-static fabric sampling and PEG enrichment technology shows higher sensitivity and stability in the etiological surveillance of large-scale object surfaces, raising the isolation rate.

     

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