Evaluation of an inactivated sample preservation solution for rapid detection of SARS-CoV-2

doi:10.3877/cma.j.issn.2095-5820.2022.01.004

Abstract

Abstract:

Objective

To evaluate the inactivation effect of sample preservation solution containing Triton X-100 and NP40 on novel coronavirus SARS-CoV-2 and its effect on rapid nucleic acid detection.

Methods

The virus inactivation effect was assessed by measuring the change of SARS-CoV-2 virus titer before and after the treatment with sample preservation solution . The influence on the minimum detection limit and precision of SARS-CoV-2 nucleic acid detection were evaluated by elution of throat swab samples containing SARS-CoV-2 simulated virus. The stability of nucleic acid of SARS-CoV-2 in the sample preservation solution was evaluated by spiking the SARS-CoV-2 virus in the solution and storing for different time.

Results

The SARS-CoV-2 virus titer decreased at least 2.76×10⁵ times after being treated by the sample preservation solution in 5 minutes. The sample preservation solution had no effect on the sensitivity of SARS-CoV-2 nucleic acid detection, and the CV of SARS-CoV-2 nucleic acid detection Ct value was less than 5%. The SARS-CoV-2 virus can be stored in the sample preservation solution for at least 4 days at 25 ℃ or 2~8 ℃.

Conclusion

The sample storage solution containing surfactants Triton X-100 and NP40 can effectively inactivate virus, and the virus nucleic acid detection is not affected after inactivation, which is suitable for rapid nucleic acid detection.

Key words: Coronaviridae, Organ preservation solutions, Surface-active agents, Nucleic acids

Li Zhao, Roujian Lu, Fei Ye, Baoying Huang, Huijuan Wang, Wenjie Tan. Evaluation of an inactivated sample preservation solution for rapid detection of SARS-CoV-2[J]. Chinese Journal of Clinical Laboratory Management(Electronic Edition), 2022, 10(01): 17-22.

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Figures/Tables 4

References 30

1	Zhu N, Zhang D, Wang W, et al. A novel coronavirus from patients with pneumonia in China, 2019[J]. N Engl J Med, 2020, 382(8): 727-733.
2	Li Q, Guan X, Wu P, et al. Early transmission dynamics in Wuhan, China, of novel coronavirus-infected pneumonia[J]. N Engl J Med, 2020, 382(13): 1199-1207.
3	Rothe C, Schunk M, Sothmann P, et al. Transmission of 2019-nCoV infection from an asymptomatic contact in Germany[J]. N Engl J Med, 2020, 382(10): 970-971.
4	Van DN, Bushmaker T, Morris DH, et al. Aerosol and surface stability of SARS-CoV-2 as compared with SARS-CoV-1[J]. N Engl J Med, 2020, 382(16): 1564-1567.
5	World Health Organization. Weekly epidemiological update on COVID-19[R]. WHO, 2021.
6	World Health Organization. Laboratory testing for coronavirus disease (COVID-19) in suspected human cases: interim guidance[R]. WHO, 2020.
7	Zachariah R, Harries AD. The WHO clinical case definition for suspected cases of Ebola virus disease arriving at Ebola holding units: reason to worry?[J]. Lancet Infect Dis, 2015, 15(9): 989-990.
8	Zhen W, Smith E, Manji R, et al. Clinical evaluation of three sample-to-answer platforms for detection of SARS-CoV-2[J]. J Clin Microbiol, 2020, 58(8): e00783-20.
9	Smithgall MC, Scherberkova I, Whittier S, et al. Comparison of Cepheid Xpert Xpress and Abbott ID now to roche cobas for the rapid detection of SARS-CoV-2[J]. J Clin Virol, 2020, 128: 104428.
10	Wolters F, Van de Bovenkamp J, Van den Bosch B, et al. Multi-center evaluation of Cepheid Xpert Xpress SARS-CoV-2 point-of-care test during the SARS-CoV-2 pandemic[J]. J Clin Virol, 2020, 128: 104426.
11	国务院应对新型冠状病毒肺炎疫情联防联控机制医疗救治组. 医疗机构新型冠状病毒核酸检测工作手册(试行第二版)[J]. 中国病毒病杂志, 2021, 11(3): 163-167.
12	马亮, 崔淑娟, 韩呈武, 等. 核酸快速检测系统在新型冠状病毒检测中的应用评价[J]. 中华预防医学杂志, 2021, 55(2): 219-225.
13	Adams NM, Leelawong M, Benton A, et al. COVID-19 diagnostics for resource-limited settings: Evaluation of "unextracted" qRT-PCR[J]. J Med Virol, 2021, 93(1): 559-563.
14	Pastorino B, Touret F, Gilles M, et al. Evaluation of heating and chemical protocols for inactivating SARS-CoV-2[J]. Viruses, 2020, 12(6): 624.
15	Loveday EK, Hain KS, Kochetkova I, et al. Effect of inactivation methods on SARS-CoV-2 virion protein and structure[J]. Viruses, 2021, 13(4): 562.
16	Jahromi R, Mogharab V, Jahromi H, et al. Synergistic effects of anionic surfactants on coronavirus (SARS-CoV-2) virucidal efficiency of sanitizing fluids to fight COVID-19[J]. Food Chem Toxicol, 2020, 145: 111702.
17	Wang T, Lien C, Liu S, et al. Effective heat inactivation of SARS-CoV-2[EB/OL]. MedRxiv, 2020. URL
18	Patterson EI, Prince T, Anderson ER, et al. Methods of inactivation of SARS-CoV-2 for downstream biological assays[J]. J Infect Dis, 2020, 222(9): 1462-1467.
19	Chan KH, Sridhar S, Zhang RR, et al. Factors affecting stability and infectivity of SARS-CoV-2[J]. J Hosp Infect, 2020, 106(2): 226-231.
20	Pere H, Podglajen I, Baillard JL, et al. Thermal inactivation and nucleic acid amplification-based testing for SARS-CoV-2[J]. J Clin Virol, 2020, 131: 104588.
21	段秀枝, 王旭楚, 俞攀, 等. 病毒灭活处理对2019新型冠状病毒核酸检测弱阳性结果的影响[J]. 中华检验医学杂志, 2020, 43(4): 358-363.
22	姜蕾, 张丽媛, 刘大宁. 两种灭活方法对2019新型冠状病毒咽拭子标本病毒核酸检测结果的影响[J]. 分子诊断与治疗杂志, 2020, 12(4): 4.
23	Yan Y, Chang L, Wang L. Laboratory testing of SARS-CoV, MERS-CoV, and SARS-CoV-2 (2019-nCoV): Current status, challenges, and countermeasures[J]. Rev Med Virol, 2020, 30(3): e2106.
24	Tawfik SM, Abd-Elaal AA, Shaban SM, et al. Surface, thermodynamic and biological activities of some synthesized Gemini quaternary ammonium salts based on polyethylene glycol[J]. J Ind Eng Chem, 2015, 30: 112-119.
25	Zhang S, Ding S, Yu J, et al. Antibacterial activity, in vitro cytotoxicity, and cell cycle arrest of gemini quaternary ammonium surfactants[J]. Langmuir, 2015, 31(44): 12161-12169.
26	Chepurnov AA, Bakulina LF, Dadaeva AA, et al. Inactivation of Ebola virus with a surfactant nanoemulsion[J]. Acta Trop, 2003, 87(3): 315-320.
27	Hamouda T, Myc A, Donovan B, et al. A novel surfactant nanoemulsion with a unique non-irritant topical antimicrobial activity against bacteria, enveloped viruses and fungi[J]. Microbiol Res, 2001, 156(1): 1-7.
28	Asculai SS, Weis MT, Rancourt MW, et al. Inactivation of herpes simplex viruses by nonionic surfactants[J]. AAC, 1978, 13(4): 686-690.
29	Patterson EI, Prince T, Anderson ER, et al. Methods of inactivation of SARS-CoV-2 for downstream biological assays[J]. JID, 2020, 222(9): 1462-1467.
30	Welch SR, Davies KA, Buczkowski H, et al. Analysis of inactivation of SARS-CoV-2 by specimen transport media, nucleic acid extraction reagents, detergents, and fixatives[J]. J Clin Microbiol, 2020, 58(11): e01713-01720.

组别	浓度（拷贝/ml）	Ct（平均）		检出率（%）
组别	浓度（拷贝/ml）	FAM	ROX	FAM	ROX
实验组	800	20.46	20.74	100	100
	400	21.86	22.02	100	95
	200	23.69	23.84	70	55
	100	23.70	24.03	25	15

对照组	800	20.08	20.91	100	100
	400	21.82	22.18	100	95
	200	23.51	23.90	65	55
	100	23.72	23.85	25	15

组别	浓度（拷贝/ml）	Ct（平均）		检出率（%）
组别	浓度（拷贝/ml）	FAM	ROX	FAM	ROX
实验组	800	20.46	20.74	100	100
	400	21.86	22.02	100	95
	200	23.69	23.84	70	55
	100	23.70	24.03	25	15

对照组	800	20.08	20.91	100	100
	400	21.82	22.18	100	95
	200	23.51	23.90	65	55
	100	23.72	23.85	25	15

项目	ORF 1ab Ct值			N基因Ct值
项目	20200726	20200933	20200934	20200726	20200933	20200934
1	18.62	19.10	19.45	18.70	18.96	18.76
2	18.73	18.71	19.06	18.44	19.46	18.90
3	17.78	19.77	19.45	18.46	18.65	18.71
4	19.56	19.82	19.13	18.73	19.06	19.38
5	18.97	19.78	19.20	18.34	18.39	18.45
6	19.60	19.82	19.23	18.92	18.81	19.54
7	19.02	19.04	19.11	17.00	18.71	19.32
8	19.22	19.36	18.69	17.87	18.25	18.61
9	19.13	18.96	18.47	18.53	18.65	18.49
10	18.47	19.15	18.97	17.19	18.46	19.07
批内CV（%）	2.87	2.15	1.61	3.60	1.89	2.05
批间CV（%）	2.39			3.00

项目	ORF 1ab Ct值			N基因Ct值
项目	20200726	20200933	20200934	20200726	20200933	20200934
1	18.62	19.10	19.45	18.70	18.96	18.76
2	18.73	18.71	19.06	18.44	19.46	18.90
3	17.78	19.77	19.45	18.46	18.65	18.71
4	19.56	19.82	19.13	18.73	19.06	19.38
5	18.97	19.78	19.20	18.34	18.39	18.45
6	19.60	19.82	19.23	18.92	18.81	19.54
7	19.02	19.04	19.11	17.00	18.71	19.32
8	19.22	19.36	18.69	17.87	18.25	18.61
9	19.13	18.96	18.47	18.53	18.65	18.49
10	18.47	19.15	18.97	17.19	18.46	19.07
批内CV（%）	2.87	2.15	1.61	3.60	1.89	2.05
批间CV（%）	2.39			3.00

项目	ORF 1ab Ct值			N基因 Ct值
项目	20200726	20200933	20200934	20200726	20200933	20200934
1	16.09	16.64	16.38	16.25	16.08	16.03
2	16.54	16.97	15.48	16.09	16.52	16.29
3	16.40	15.28	16.90	16.27	16.11	16.72
4	16.40	15.10	16.60	16.01	15.97	16.04
5	16.49	15.29	16.88	16.31	16.93	16.53
6	16.40	15.28	16.88	16.77	16.16	16.04
7	16.65	15.75	16.13	15.98	16.31	16.16
8	16.39	15.55	15.87	15.62	15.85	15.79
9	16.54	15.38	16.23	15.98	15.13	16.23
10	16.53	15.83	15.75	15.60	16.40	16.14
批内CV（%）	0.92	3.98	3.11	2.13	2.92	1.65
批间CV（%）	3.47			2.23

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