Comparative analysis of nucleic acid PCR orthogonal experiment of first monkeypox virus sample in mainland China

Hua ZONG; Qiao SUN; Caiyun LI; Jing YIN; Qingmei LUO; Mengjia WANG

doi:10.16016/j.2097-0927.202210035

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Comparative analysis of nucleic acid PCR orthogonal experiment of first monkeypox virus sample in mainland China

Public Health and Preventive Medicine | 更新时间：2026-01-09

- Comparative analysis of nucleic acid PCR orthogonal experiment of first monkeypox virus sample in mainland China
- Journal of Army Medical University Vol. 44, Issue 23, Pages: 2440-封3(2022)
- 作者机构：
  
  1.400067 重庆，重庆市南岸区疾病预防控制中心微生物检验科
  2.400016 重庆，重庆医科大学公共卫生学院
- 作者简介：
- 基金信息：
- DOI：10.16016/j.2097-0927.202210035
  CLC： R195.1;R373.12;R446.61
- Received：08 October 2022，
  
  Revised：2022-10-21，
  
  Published：15 December 2022
- 稿件说明：
移动端阅览
Hua ZONG, Qiao SUN, Caiyun LI, et al. Comparative analysis of nucleic acid PCR orthogonal experiment of first monkeypox virus sample in mainland China[J]. Journal of Army Medical University, 2022, 44(23): 2440-封3.
DOI：

Hua ZONG, Qiao SUN, Caiyun LI, et al. Comparative analysis of nucleic acid PCR orthogonal experiment of first monkeypox virus sample in mainland China[J]. Journal of Army Medical University, 2022, 44(23): 2440-封3. DOI： 10.16016/j.2097-0927.202210035.

摘要

目的

验证目前国内关于猴痘病毒提取试剂、扩增试剂以及实时荧光定量聚合酶链式反应(real-time quantitativepolymerase chain reaction，RT-qPCR)仪的检出效率、灵敏度差异以及提取扩增组合的比对分析。

方法

根据设计的正交试验方案，采用RT-qPCR对提取的核酸进行扩增。

结果

本次试验中不同的DNA提取试剂的提取效率的差异无统计学意义(

=12.855，

=0.070)，扩增试剂扩增循环数的差异(

=49.832，

=0.019)和RT-qPCR仪对荧光捕捉信号的差异(

=36.355，

=0.027)均存在统计学意义。

结论

目前国内猴痘病毒的提取和扩增试剂盒可以有效地检出该病毒，试剂和仪器的组合对检出效率存在差异。通过验证试验，发现更具有检测优势的组合，为之后国内应对猴痘病毒检测及疫情防控工作提供参考。

Abstract

Objective

To investigate the differences in detection efficiency and sensitivity of current domestic monkeypox virus extraction reagents

amplification reagents and real-time quantitative polymerase chain reaction (RT-qPCR) instruments

and to compare among different extraction and amplification combinations.

Methods

The extracted nucleic acid of monkeypox virus were amplified using RT-qPCR

according to the designed orthogonal test protocol.

Results

There was no significance in the extraction efficiency of different DNA extraction reagents (

=12.855

=0.07)

while the differences were statistically differences in the number of amplification cycles of amplification reagents (

=49.832

=0.019) and in the fluorescence signal capture of RT-qPCR instruments (

=36.355

=0.027).

Conclusion

The current domestic extraction and amplification kits for monkeypox virus can effectively detect the virus

but various reagent and instrument combinations show different detection efficiency. Through the validation test

a more advantageous combination is found

which provides a reference for domestic response to monkeypox virus detection and epidemic prevention and control.

关键词

Keywords

references

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