摘要
收集11种不同厂家的灭活型样本保存液,将甲型流感病毒加入保存液和生理盐水中,在4℃、25℃和37℃条件下保存0 h、2 h、4 h、6 h和24 h后,分别提取各组的病毒核酸,并通过逆转录实时荧光PCR检测病毒载量变化,以探究不同样本保存液对病毒核酸保护效果是否存在差异。结果发现不同样本保存液中的病毒载量随保存时间和温度变化存在显著差异。据此可将11种灭活型样本保存液的保存效果分为四类:(1)优秀产品:在任一温度(4℃、25℃、37℃)条件下,即使保存24h时病毒核酸都未发生明显降解,占比27.2%(3/11);(2)良好产品:低温(4℃)和室温(25℃)情况下核酸未出现降解,但在高温(37℃)条件下6 h后保存效果明显下降,占比27.2%(3/11);(3)合格产品:低温(4℃)条件下对病毒核酸具有较好的保护效果,但在室温(25℃)和高温(37℃)情况下随保存时间延长其保护效果显著下降,占比18.1%(2/11);(4)不合格产品:任一保存条件下对病毒核酸的保护效果都较差,甚至加入病毒后立即导致病毒核酸发生快速降解,占比27.2%(3/11)。以上结果说明不同厂家样本保存液对病毒核酸的保护效果存在显著差异,可能会导致病毒核酸检测出现假阴性结果。因此,需重点加强对样本保存液的质量监管,以有效保障病毒核酸检测,尤其是新冠核酸检测质量,以更好地做好疫情防控工作。
Inactivated sample-preservation solutions(SPSs) from 11 manufacturers were collected. Influenza A virus was added to each SPS and saline. Viral nucleic acids was extracted from each group after being storage at 4 ° C, 25 ° C and 37 ° C for 0, 2, 4, 6 and 24 h. Changes in viral loads were detected by real-time reverse transcription fluorescence polymerase chain reaction to identify differences in the protection of VNAs in different SPSs. We discovered that the viral load in different SPSs varied significantly with the preservation time and temperature. The 11 inactivated SPSs could be classified into four categories. The first category was “excellent products”(VNAs were not degraded significantly at any temperature(4°C, 25°C, 37°C) even after storage for 24 h) and accounted for 27.2%(3/11). The second category was “good products”(no degradation of VNAs occurred at low temperature(4° C) and room temperature(25° C), but the preservation effect decreased significantly after 6 h at high temperature(37°C)), and accounted for 27.2%(3/11). The third category was “qualified products”(the protection of VNAs was good at low temperature(4°C), but decreased significantly with an extension of storage time at room temperature(25°C) and high temperature(37°C)) and accounted for 18.1%(2/11). The fourth category was “substandard products”(protection of VNAs was poor under either storage condition, and even caused rapid degradation of VNAs immediately after the addition of virus) and accounted for 27.2%(3/11). Our results indicated a significant difference in the effect of SPSs from different manufacturers to protect VNAs from degradation, which could lead to false-negative results in VNA testing.Therefore, strengthening the quality control of SPSs must be done to guarantee the quality of VNA testing for better prevention and control of epidemics.
作者
史春丽
黄中强
范晓瑜
肖艳群
王雪亮
SHI Chunli;HUANG Zhongqiang;FAN Xiaoyu;XIAO Yanqun;WANG Xueliang(Department of Molecular Biology Shanghai Center for Clinical Laboratory,Shanghai 200126,China)
出处
《病毒学报》
CAS
CSCD
北大核心
2023年第2期477-483,共7页
Chinese Journal of Virology
基金
上海市公共卫生体系建设三年行动计划(项目号:GWV-2)
上海市“医苑新星”青年医学人才培养资助计划(青年医学人才类-临床检验项目)
上海市卫生健康委员会面上项目(项目号:201940264),题目:液态活检中热点突变循环肿瘤DNA参考物质的制备与应用
上海市临床检验中心自选课题(项目号:2022ZXKT-03),题目:11种灭活型病毒保存液对病毒核酸稳定性及其灭活效果的影响研究。
关键词
样本保存液
保护效果
甲型流感病毒
逆转录实时荧光PCR
新型冠状病毒
Sample-preservation solution
Protective effect
Influenza A virus
Real-time reverse transcription fluorescent PCR
Coronavirus disease 2019(COVID-19)