Establishment of rapid detection method for zika virus based on direct amplification RT-PCR technique
10.3969/j.issn.1673-4130.2024.03.019
- VernacularTitle:基于直扩RT-PCR技术的寨卡病毒快速检测方法的建立
- Author:
Lang LI
1
,
2
;
Libing GU
;
Li ZHU
;
Jianan HE
;
Ying YE
;
Ran ZHANG
;
Huawen LI
;
Fuyuan LI
;
Dayong GU
Author Information
1. 广东医科大学公共卫生学院,广东东莞 523808
2. 深圳市检验检疫科学研究院,广东深圳 518033
- Keywords:
zika virus;
direct amplification real-time fluorescent quantitative reverse transcription poly-merase chain reaction technique;
DNA polymerase
- From:
International Journal of Laboratory Medicine
2024;45(3):358-364
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a rapid detection method for zika virus based on direct amplification re-al-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR)technique.Methods A direct amplification RT-PCR technique for the rapid detection of zika virus in 5 samples(whole blood,serum,saliva,throat swab and urine)was established by using a special function DNA polymerase and a preferred PCR enhancer.Results The detection limits of the 5 samples were 103 PFU/mL in serum,102 PFU/mL in urine,throat swab,and saliva,and 104 PFU/mL in whole blood.The coefficient of goodness-fit of stand-ard curves was above 0.98,and the amplification efficiency was 90％-110％.Zika virus nucleic acid was suc-cessfully amplified,but non-zika virus nucleic acid was not amplified.Based on the repeatable detection of sam-ples from urine,whole blood,and saliva,the variation coefficient of 6 repeated Ct values at 106 PFU/mL and 102 PFU/mL concentrations were all＜5％.The zika virus detection method established by the direct amplifi-cation RT-PCR technique was consistent with the detection results of conventional RT-PCR technique.Only two serum samples were detected in eight zika virus samples,and the remaining 62 non-zika virus samples and 12 negative samples were not amplified.Conclusion A rapid detection method for zika virus based on direct ampli-fication RT-PCR technique is successfully established.The method is simple,rapid,sensitive and specific.
