Use of guanidine thiocyanate-based nucleic acid extraction buffers to inactivate poliovirus in potentially infectious materials

Honeywood, Michelle J.; Jeffries-Miles, Stacey; Wong, Kimberly; Harrington, Chelsea; Burns, Cara C.; Oberste, M. Steven; Bowen, Michael D.; Vega, Everardo

doi:10.1016/j.jviromet.2021.114262

Cited by 8 publications

(6 citation statements)

References 22 publications

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“…As a result, it is commonly included in nucleic acid extraction buffers. 23 For example, TRIzol reagent was a single-phase solution containing phenol and GITC, which is often used for deproteinization to extract RNA. 24 Since the samples were unable to meet the required refrig- However, the advantage of the study of pseudovirus particles is that it can make a standardized curve to accurately quantify the degree of degradation of the SARS-CoV-2 pseudovirus particles so that the degree of degradation can be expressed in a digital form.…”

Section: Discussionmentioning

confidence: 99%

“…Previous research has demonstrated that GIHCl exhibits a superior ability to effectively eradicate viruses and inhibit RNase. As a result, it is commonly included in nucleic acid extraction buffers 23 . For example, TRIzol reagent was a single‐phase solution containing phenol and GITC, which is often used for deproteinization to extract RNA 24 …”

Section: Discussionmentioning

confidence: 99%

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How to better select SARS‐CoV‐2 preservation solution of virus nucleic acid testing

Duan,

Huang,

Wang

et al. 2023

Clinical Laboratory Analysis

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BackgroundSampling and testing for SARS‐CoV‐2 is a widely recognized method for identifying patients with COVID‐19. However, there is limited research available on the stability of nucleic acids in viral storage solutions.MethodsThis paper investigates the components that provide better protection for virus and nucleic acid detection. The study utilized real‐time quantitative fluorescent PCR to detect SARS‐CoV‐2 and evaluate the preservation effect and stability of SARS‐CoV‐2 viral storage solution under various conditions, including different guanidinium salts, brands, and storage conditions.ResultsAll brands of inactivated virus preservation solutions demonstrated effective preservation and stability. However, 0.5 mol/L guanidine hydrochloride and guanidine isothiocyanate solutions exhibited poor antiseptic effects. Additionally, refrigerated storage showed better preservation compared to room temperature storage.ConclusionsWe recommend using inactivated virus collection solution to preserve and transport samples and testing preferably within 6 hours to reduce false negatives of NAT results.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

How to better select SARS‐CoV‐2 preservation solution of virus nucleic acid testing

Duan,

Huang,

Wang

et al. 2023

Clinical Laboratory Analysis

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“…The optimized protocol for kit B was simpler and the reaction was faster than those of kit A; with the addition of all the primers at the start of reaction, the RT-PCR reaction finished within 2.5 h; however, the efficient amplification could negatively affect the detection rate of minor populations of PV strains in the mixtures ( Table 2 , see below discussion). Selection of commercially available kits is apparently a critical factor for the performance of the DD, including the extraction step of viral RNA [ 25 , 26 ].…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Direct Detection Protocols for Poliovirus from Stool Samples of Acute Flaccid Paralysis Patients

Ueno,

Kitamura,

Nishimura

et al. 2023

Viruses

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Polio surveillance in the Global Polio Eradication Initiative has been conducted with virus isolation from stool samples of acute flaccid paralysis (AFP) cases. Under the current biorisk management/regulations, challenges arise in the timelines of the report, sensitivity of the test and containment of poliovirus (PV) isolates. In the present study, we evaluated protocols of previously reported direct detection (DD) methods targeting the VP1 or VP4–VP2 regions of the PV genome in terms of sensitivity and sequencability. An optimized protocol targeting the entire-capsid region for the VP1 sequencing showed a high sensitivity (limit of detection = 82 copies of PV genome) with a simpler and faster reaction than reported ones (i.e., with the addition of all the primers at the start of the reaction, the RT-PCR reaction finishes within 2.5 h). The DD methods targeting the VP1 region detected PV in 60 to 80% of PV-positive stool samples from AFP cases; however, minor populations of PV strains in the samples with virus mixtures were missed by the methods. Sequencability of the DD methods was primarily determined by the efficiency of the PCRs for both Sanger and nanopore sequencing. The DD method targeting the VP4–VP2 region showed higher sensitivity than that targeting the VP1 region (limit of detection = 25 copies of PV genome) and successfully detected PV from all the stool samples examined. These results suggest that DD methods are effective for the detection of PV and that further improvement of the sensitivity is essential to serve as an alternative to the current polio surveillance algorithm.

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“…As indicated by the publicly available safety data sheet, this reagent contains a mixture of guanidine thiocyanate, ethanol, and n-lauroylsarcosine ( 19 ). Guanidine thiocyanate-based reagents have previously been shown to effectively inactivate poliovirus and FMDV, though the comparison of results should be interpreted with caution as the exact formulation of PSMTM is proprietary ( 20 , 21 ). Previous studies have shown that ethanol and n-lauroylsarcosine inactivate various enveloped and some non-enveloped viruses by interacting with lipids and denaturing proteins ( 22 ).…”

Section: Introductionmentioning

confidence: 99%

Inactivation of highly transmissible livestock and avian viruses including influenza A and Newcastle disease virus for molecular diagnostics

Welch,

Shrestha,

Hutchings

et al. 2024

Front. Vet. Sci.

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There is a critical need for an inactivation method that completely inactivates pathogens at the time of sample collection while maintaining the nucleic acid quality required for diagnostic PCR testing. This inactivation method is required to alleviate concerns about transmission potential, minimize shipping complications and cost, and enable testing in lower containment laboratories, thereby enhancing disease diagnostics through improved turn-around time. This study evaluated a panel of 10 surrogate viruses that represent highly pathogenic animal diseases. These results showed that a commercial PrimeStore® molecular transport media (PSMTM) completely inactivated all viruses tested by >99.99%, as determined by infectivity and serial passage assays. However, the detection of viral nucleic acid by qRT-PCR was comparable in PSMTM and control-treated conditions. These results were consistent when viruses were evaluated in the presence of biological material such as sera and cloacal swabs to mimic diagnostic sample conditions for non-avian and avian viruses, respectively. The results of this study may be utilized by diagnostic testing laboratories for highly pathogenic agents affecting animal and human populations. These results may be used to revise guidance for select agent diagnostic testing and the shipment of infectious substances.

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Use of guanidine thiocyanate-based nucleic acid extraction buffers to inactivate poliovirus in potentially infectious materials

Cited by 8 publications

References 22 publications

How to better select SARS‐CoV‐2 preservation solution of virus nucleic acid testing

How to better select SARS‐CoV‐2 preservation solution of virus nucleic acid testing

Evaluation of Direct Detection Protocols for Poliovirus from Stool Samples of Acute Flaccid Paralysis Patients

Inactivation of highly transmissible livestock and avian viruses including influenza A and Newcastle disease virus for molecular diagnostics

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