Rapid, sensitive, and specific, low-resource molecular detection of Hendra virus

Pollak, Nina M.; Marsh, Glenn A.; Olsson, Malin A; McMillan, David; Macdonald, Joanne

doi:10.1016/j.onehlt.2023.100504

Cited by 7 publications

(8 citation statements)

References 36 publications

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“…However, the use of a chemical inactivation and extraction process, such as TNA-Cifer Reagent E, is not likely to be significantly influenced by amino acid changes in the viral proteins. Although the inactivation of the Hendra virus, Nipah virus, and dengue virus has been demonstrated (Pollak et al, 2022b , 2023a , b ), the inactivation of common human pathogens, such as HIV and hepatitis viruses, has not yet been demonstrated, with such viruses being an established safety concern for blood tests. The influence of VTM and swabs on the ability of TNA-Cifer Reagent E to inactivate SARS-CoV-2 was also not evaluated, with SARS-CoV-2 inactivation potentially influenced by different excipients and their interactions with various patient-derived materials.…”

Section: Discussionmentioning

confidence: 99%

“…A simple new sample preparation reagent, TNA-Cifer Reagent E, has been described for use in RT-qPCR-based detection in a number of infectious disease settings (Ahmed et al, 2022a , b ; Pollak et al, 2022a , b , 2023a , b ). In this study, we describe a pre-clinical evaluation of the use of this reagent for the RT-qPCR-based detection of SARS-CoV-2.…”

Section: Introductionmentioning

confidence: 99%

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Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E

Pollak,

Rawle,

Yan

et al. 2023

Front. Microbiol.

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RT-qPCR remains a key diagnostic methodology for COVID-19/SARS-CoV-2. Typically, nasal or saliva swabs from patients are placed in virus transport media (VTM), RNA is extracted at the pathology laboratory, and viral RNA is measured using RT-qPCR. In this study, we describe the use of TNA-Cifer Reagent E in a pre-clinical evaluation study to inactivate SARS-CoV-2 as well as prepare samples for RT-qPCR. Adding 1 part TNA-Cifer Reagent E to 5 parts medium containing SARS-CoV-2 for 10 min at room temperature inactivated the virus and permitted RT-qPCR detection. TNA-Cifer Reagent E was compared with established column-based RNA extraction and purification methodology using a panel of human clinical nasal swab samples (n = 61), with TNA-Cifer Reagent E showing high specificity (100%) and sensitivity (97.37%). Mixtures of SARS-CoV-2 virus and TNA-Cifer Reagent E could be stored for 3 days at room temperature or for 2 weeks at 4°C without the loss of RT-qPCR detection sensitivity. The detection sensitivity was preserved when TNA-Cifer Reagent E was used in conjunction with a range of VTM for saliva samples but only PBS (Gibco) and Amies Orange for nasal samples. Thus, TNA-Cifer Reagent E improves safety by rapidly inactivating the virus during sample processing, potentially providing a safe means for molecular SARS-CoV-2 testing outside traditional laboratory settings. The reagent also eliminates the need for column-based and/or automated viral RNA extraction/purification processes, thereby providing cost savings for equipment and reagents, as well as reducing processing and handling times.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E

Pollak,

Rawle,

Yan

et al. 2023

Front. Microbiol.

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“…ZIKV MR766 culture (accession MK105975) was used to infect C6/36 cells at a multiplicity of Infection (MOI) of 0.01 for 5 and 7 days, respectively ( Pollak et al, 2023a ). Virus culture media was harvested after centrifugation at 4°C for 10 min at 130 × g and then stored at −80°C.…”

Section: Methodsmentioning

confidence: 99%

Development and pre-clinical evaluation of a Zika virus diagnostic for low resource settings

Balea,

Pollak,

Hobson-Peters

et al. 2023

Front. Microbiol.

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IntroductionZika virus (ZIKV) is a re-emerging flavivirus that poses a significant public health threat. ZIKV exhibits a wide array of non-vector borne human transmission routes, such as sexual transmission, transplacental transmission and blood transfusion. Detection and surveillance of ZIKV is considered paramount in prevention of major outbreaks. With the majority of cases reported in low-resource locations, simple, low-cost detection methods are considered highly desirable.Materials and MethodsHere we have developed a sensitive and specific ZIKV diagnostic using reverse transcription recombinase-aided amplification (RT-RAA) coupled with lateral flow detection (LFD) targeting a highly conserved region of the ZIKV NS1 gene.ResultsWe show our rapid, isothermal-ZIKV-diagnostic (Iso-ZIKV-Dx) can detect 500 copies of synthetic ZIKV RNA/μL in under 30 min at a constant 39°C. Using simulated urine samples, we observed that Iso-ZIKV-Dx also detects as low as 34.28 RNA copies/reaction of ZIKV (MR766 strain). Specificity testing confirmed that our test does not detect any co-circulating flaviviruses (dengue, West Nile, Japanese encephalitis, Murray Valley encephalitis and yellow fever viruses) or chikungunya virus. Sample processing results show complete inactivation of ZIKV (MR766 strain) in 5 min at room temperature using our novel viral RNA sample preparation reagent. Furthermore, lateral flow strips testing demonstrates positive diagnoses in as little as 5 min in running buffer.DiscussionContrary to conventional RT-qPCR, our Iso-ZIKV-Dx does not require expensive machinery, specialised laboratory settings or extensively trained personnel. Pre-clinical evaluation demonstrates that our test exhibits robust, in-field capabilities without compromising sensitivity or specificity. When compared to the gold-standard RT-qPCR, our Iso-ZIKV-Dx test offers an array of applications that extend beyond diagnostics alone, including potential for surveillance and monitoring of ZIKV vector competency.

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“…Specific Nipah virus PCR assays are used to test clinical samples, such as blood, urine, throat swabs, or cerebrospinal fluid, that are obtained from suspected cases. Since PCR has such high sensitivity and specificity, it's frequently chosen for early diagnosis [45].…”

Section: Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Nipah Virus Disease: An Updated Review

Vandana,

Swarna,

Sreevidya

et al. 2024

UPJOZ

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The Nipah virus (NiV) is a member of the Paramyxoviridae family of zoonotic viruses that is extremely contagious and potentially fatal. The Nipah virus was first discovered in 1999 during an outbreak in Malaysia. Since then, it has periodically caused outbreaks throughout South and Southeast Asia, especially in Bangladesh and India, and it continues to be a serious public health concern. The virus is primarily transmitted to humans through direct contact with infected fruit bats, which serve as natural reservoir hosts for the Nipah virus. Human-to-human transmission can also happen when infected people's body fluids come into close contact with one another. The Nipah virus has a high death rate that varies depending on the outbreak and can cause a variety of clinical presentations, including encephalitis, severe respiratory illnesses, and asymptomatic infections. Fever, headache, dizziness, drowsiness, and confusion are common signs of Nipah virus infections, which may lead to the rapid onset of coma in the patient. The Nipah virus does not presently have a specific antiviral treatment; instead, infection control methods and supportive care are the foundation of management to stop the virus from spreading. Nipah virus outbreaks must be stopped with an approach that includes community involvement, public health education, and surveillance of both human and animal populations. Additionally, efforts are directed toward developing vaccines as a more potent form of prevention. The Nipah virus is a clear reminder of the continuous threat that newly emerging infectious diseases pose, as well as the necessity of having strong surveillance systems and quick response times in place to lessen the effects that these outbreaks have on public health.

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Rapid, sensitive, and specific, low-resource molecular detection of Hendra virus

Cited by 7 publications

References 36 publications

Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E

Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E

Development and pre-clinical evaluation of a Zika virus diagnostic for low resource settings

Nipah Virus Disease: An Updated Review

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