Reality Check of Laboratory Service Effectiveness during Pandemic (H1N1) 2009, Victoria, Australia

Catton, Michael; Druce, Julian; Papadakis, Georgina; Tran, Thomas; Birch, Christopher

doi:10.3201/eid/1706.101747

Cited by 5 publications

(2 citation statements)

References 5 publications

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“…Reduced reaction volumes had previously been validated (36). RT was performed using the following conditions: 25°C for 10 min, 42°C for 15 min, 85°C for 5 min, and a 4°C hold.…”

Section: Methodsmentioning

confidence: 99%

Neutralization Assay for Zika and Dengue Viruses by Use of Real-Time-PCR-Based Endpoint Assessment

et al. 2017

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The global spread and infective complications of Zika virus (ZKV) and dengue virus (DENV) have made them flaviviruses of public health concern. Serological diagnosis can be challenging due to antibody cross-reactivity, particularly in secondary flavivirus infections or when there is a history of flavivirus vaccination. The virus neutralization assay is considered to be the most specific assay for measurement of anti-flavivirus antibodies. This study describes an assay where the neutralization endpoint is measured by real-time PCR, providing results within 72 h. It demonstrated 100% sensitivity (24/24 ZKV and 15/15 DENV) and 100% specificity (11/11 specimens) when testing well-characterized sera. In addition, the assay was able to determine the correct DENV serotype in 91.7% of cases. The high sensitivity and specificity of the real-time PCR neutralization assay makes it suitable to use as a confirmatory test for sera that are reactive in commercial IgM/IgG enzyme immunoassays. Results are objective and the PCR-based measurement of the neutralization endpoint lends itself to automation so that throughput may be increased in times of high demand.KEYWORDS dengue, flavivirus, serology, Zika Z ika virus (ZKV) and dengue virus (DENV) are members of the family Flaviviridae, genus Flavivirus, which also includes yellow fever, Japanese encephalitis, and West Nile viruses. There are at least 4 closely related but serologically distinct DENV, designated DENV serotypes 1 through 4 (1). There is only transient and weak crossprotection among the 4 DENV serotypes, so individuals living in an area where DENV is endemic can be infected up to 4 times (2).ZKV and DENV have spread across the tropics in Africa, Asia, the Pacific, and the Americas with their vector mosquitoes Aedes aegypti and A. albopictus (3, 4). Cocirculation of ZKV and DENV types 1 through 4 has been documented in both French Polynesia and Brazil (5, 6) and likely occurs in many other tropical countries. Incursions into temperate countries have also occurred, with autochthonous transmission observed in Europe (DENV in southeastern France, Croatia, and Madeira Island) and the United States (DENV and ZKV in Florida) (7-9).Both ZKV and DENV infection can produce a wide spectrum of illnesses with many asymptomatic or subclinical infections (10, 11). ZKV's potential to cause fetal central nervous system abnormalities, growth restriction, and death (12) has necessitated ZKV testing of potentially exposed pregnant women, among whom nonspecific serological activity may occur, and asymptomatic individuals with low pretest probabilities of positive ZKV serology.Direct detection of ZKV nucleic acids has limited utility in asymptomatic patients, as the periods of viremia and shedding in urine and saliva are relatively brief, occurring

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“…Reduced reaction volumes had previously been validated (36). RT was performed using the following conditions: 25°C for 10 min, 42°C for 15 min, 85°C for 5 min, and a 4°C hold.…”

Section: Methodsmentioning

confidence: 99%

Neutralization Assay for Zika and Dengue Viruses by Use of Real-Time-PCR-Based Endpoint Assessment

et al. 2017

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“… 1 , 2 Recent studies have demonstrated variability in current shipping conditions both in terms of expediency and temperature exposure of the sample. 3 , 4 From these studies, one may conclude that sample stabilization prior to transport may be crucial to ensure accuracy of downstream diagnostic testing. To facilitate sample collection and storage in austere environments, such as those experienced during outbreaks of tropical diseases, a stabilization product ideally needs to be inexpensive as well as easy to use, requiring minimal ancillary equipment.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Nucleic Acid Stabilization Products for Ambient Temperature Shipping and Storage of Viral RNA and Antibody in a Dried Whole Blood Format

Dauner¹,

Gilliland²,

Mitra³

et al. 2015

The American Society of Tropical Medicine and Hygiene

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Loss of sample integrity during specimen transport can lead to false-negative diagnostic results. In an effort to improve upon the status quo, we used dengue as a model RNA virus to evaluate the stabilization of RNA and antibodies in three commercially available sample stabilization products: Whatman FTA Micro Cards (GE Healthcare Life Sciences, Pittsburgh, PA), DNAstāble Blood tubes (Biomātrica, San Diego, CA), and ViveST tubes (ViveBio, Alpharetta, GA). Both contrived and clinical dengue-positive specimens were stored on these products at ambient temperature or 37°C for up to 1 month. Antibody and viral RNA levels were measured by enzyme-linked immunosorbent assay (ELISA) and quantitative reverse transcription polymerase chain reaction (qRT-PCR) assays, respectively, and compared with frozen unloaded controls. We observed reduced RNA and antibody levels between stabilized contrived samples and frozen controls at our earliest time point, and this was particularly pronounced for the FTA cards. However, despite some time and temperature dependent loss, a 94.6–97.3% agreement was observed between stabilized clinical specimens and their frozen controls for all products. Additional considerations such as cost, sample volume, matrix, and ease of use should inform any decision to incorporate sample stabilization products into a diagnostic testing workflow. We conclude that DNAstāble Blood and ViveST tubes are useful alternatives to traditional filter paper for ambient temperature shipment of clinical specimens for downstream molecular and serological testing.

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Evaluation of Swabs, Transport Media, and Specimen Transport Conditions for Optimal Detection of Viruses by PCR

et al. 2012

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Depletion of swabs and viral transport medium during epidemics may prompt the use of unvalidated alternatives. Swabs collected and transported dry or in saline were compared to commercially available swab/medium combinations for PCR detection of influenza, enterovirus, herpes simplex virus, and adenovirus. Each was detected at an ambient temperature (22°C) and 4°C for 7 days. Detection of influenza on dry or saline swabs is important because of its capacity to cause outbreaks involving large numbers of cases.

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Reality Check of Laboratory Service Effectiveness during Pandemic (H1N1) 2009, Victoria, Australia

Abstract: TOC summary: The greatest challenges were insufficient staff and test reagents.

Cited by 5 publications

References 5 publications

Neutralization Assay for Zika and Dengue Viruses by Use of Real-Time-PCR-Based Endpoint Assessment

Neutralization Assay for Zika and Dengue Viruses by Use of Real-Time-PCR-Based Endpoint Assessment

Evaluation of Nucleic Acid Stabilization Products for Ambient Temperature Shipping and Storage of Viral RNA and Antibody in a Dried Whole Blood Format

Evaluation of Swabs, Transport Media, and Specimen Transport Conditions for Optimal Detection of Viruses by PCR

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