摘要
目的确定上海捷诺生物科技有限公司研制生产的新型冠状病毒2019-nCoV核酸检测试剂盒(荧光PCR法)最低检测限,并验证其在最低检测限水平是否可有效检出新型冠状病毒。方法将20份不同临床阳性样本稀释至62500、12500、2500、500、100 copies/mL 5个浓度梯度,利用新型冠状病毒2019-nCoV核酸检测试剂盒(荧光PCR法)进行3次重复检测,初步确定最低检测限范围,用3批生产的试剂盒对稀释至1500、700、500、300、150 copies/mL 5个浓度梯度的20份不同临床阳性样本进行20次重复检测,进一步确定最低检测限,另取25份不同临床阳性样本稀释至最低检测限水平,用连续3批生产的试剂盒进行20次重复检测,以验证其最低检测限。结果在62500、12500、2500、500 copies/mL 4个浓度水平上,样本阳性检出率均为100%,在100 copies/mL浓度水平上,样本阳性检出率≤55%,初步最低检测限在500~100 copies/mL之间,经20次重复测定,1500、700、500 copies/mL 3个浓度水平对样品的阳性检出率≥95%,300、150 copies/mL 2个浓度水平对样品的阳性检出率均<95%,确定最低检测限为500 copies/mL,25份稀释至最低检测限浓度水平的阳性样品,阳性检出率均达95%以上。结论上海捷诺生物科技有限公司研制生产的新型冠状病毒2019-nCoV核酸检测试剂盒(荧光PCR法)的最低检测限确定为500 copies/mL,经验证,该试剂盒对最低检测限水平的病毒样本可有效检出病毒核酸,符合我国对检测试剂最低检测限的相关要求。
Objective To determine the minimum detection limit of the detection kit for nucleic acid of 2019 new coronavirus(2019-nCoV)(fluorescent PCR),manufactured by Shanghai GeneoDx Biotech Co.,LTD.,and verify whether2019-nCoV at the minimum detection limit level may be detected by the kit. Methods By setting up five concentration gradients of 62 500,12 500,2 500,500 and 100 copies/mL for three repeat tests on 20 different clinically positive samples,the minimum detection limit range was preliminarily determined. Three consecutive batches of detection kit were used for determination of 20 different samples at five concentration gradients of 1 500,700,500,300 and 150 copies/mL for 20 times to further determine the minimum detection limit. Another 25 different clinical positive samples were diluted to the concentrations at minimum detection limit level and determined by three consecutive batches of kits for 20 times to verify minimum detection limit. Results All the positive detection rates of samples at concentrations of 62 500,12 500,2 500 and 500 copies/mL were 100%,while that at a concentration of 100 copies/mL was not more than 55%,and the minimum detection limit was preliminarily determined as 100~500 copies/mL. Twenty repeat tests showed that the positive detection rates of sample at concentrations of 1 500,700 and 500 copies/mL were not less than 95%,while those at 300 and 150 copies/mL were less than 95%,and the minimum detection limit was determined as 500 copies/mL.However,all the positive detection rates of 25 positive samples diluted to the concentration at minimum detection limit level were more than 95%. Conclusion The minimum detection limit of detection kit for nucleic acid of 2019-nCoV(fluorescent PCR)manufactured by Shanghai GeneoDx Biotech Co.,LTD. was determined as 500 copies/mL. It has been verified that the virus samples at the concentration of minimum detection limit could be detected by the kit,which meets the relevant national requirements for the minimum detection limit of detection reagents.
作者
董江锴
黄青红
范娟
黄迎燕
张云涛
杨晓明
DONG Jiang-kai;HUANG Qing-hong;FAN Juan;HUANG Ying-yan;ZHANG Yun-tao;YANG Xiao-ming(Shanghai GeneoDx Biotech Co.,LTD.,Shanghai 201700,China)
出处
《中国生物制品学杂志》
CAS
CSCD
北大核心
2021年第4期410-414,共5页
Chinese Journal of Biologicals
基金
“科技助力经济2020”重点专项(沪科[2020]183号)。
关键词
新型冠状病毒
核酸检测试剂盒
荧光PCR法
最低检测限
2019 New coronavirus(2019-nCoV)
Nucleic acid detection kit
Fluorescent PCR
Minimum detection limit
