Field detection of avian influenza virus in wild birds: Evaluation of a portable rRT-PCR system and freeze-dried reagents

Takekawa, John Y.; Iverson, Samuel A.; Schultz, Annie K.; Hill, Nichola J.; Cardona, Carol J.; Boyce, Walter M.; Dudley, Joseph P.

doi:10.1016/j.jviromet.2010.02.029

Cited by 23 publications

(15 citation statements)

References 25 publications

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“…In contrast, PCR‐based assays such as RRT‐PCR assays are highly sensitive (>95%) and specific and therefore widely used for routine laboratory‐based detection of FMDV. Over the last decade, a number of attempts were made to transfer the laboratory‐based RRT‐PCR assays to portable PCR equipment that can be operated in the field (Callahan et al., ; Hearps et al., ; Risatti et al., ; Tomlinson et al., ; Rasmussen et al., ; Takekawa et al., ; Madi et al., ; Molsa et al., , ; Liu et al., ; Howson et al., ). These portable PCR instruments, however, depend on precision thermocycling, are highly sophisticated and therefore expensive and delicate for field use.…”

Section: Discussionmentioning

confidence: 99%

Field-Deployable Reverse Transcription-Insulated Isothermal PCR (RT-iiPCR) Assay for Rapid and Sensitive Detection of Foot-and-Mouth Disease Virus

Ambagala

Fisher

Goolia

et al. 2016

Transbound Emerg Dis

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Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals, which can decimate the livestock industry and economy of countries previously free of this disease. Rapid detection of foot-and-mouth disease virus (FMDV) is critical to containing an FMD outbreak. Availability of a rapid, highly sensitive and specific, yet simple and field-deployable assay would support local decision-making during an FMDV outbreak. Here we report validation of a novel reverse transcription-insulated isothermal PCR (RT-iiPCR) assay that can be performed on a commercially available, compact and portable POCKIT analyser that automatically analyses data and displays '+' or '-' results. The FMDV RT-iiPCR assay targets the 3D region of the FMDV genome and was capable of detecting 9 copies of in vitro-transcribed RNA standard with 95% confidence. It accurately identified 63 FMDV strains belonging to all seven serotypes and showed no cross-reactivity with viruses causing similar clinical diseases in cloven-hoofed animals. The assay was able to identify FMDV RNA in multiple sample types including oral, nasal and lesion swabs, epithelial tissue suspensions, vesicular and oral fluid samples, even before the appearance of clinical signs. Clinical sensitivity of the assay was comparable or slightly higher than the laboratory-based real-time RT-PCR assay in use. The assay was able to detect FMDV RNA in vesicular fluid samples without nucleic acid extraction. For RNA extraction from more complex sample types, a commercially available taco mini transportable magnetic bead-based, automated extraction system was used. This assay provides a potentially useful field-deployable diagnostic tool for rapid detection of FMDV in an outbreak in FMD-free countries or for routine diagnostics in endemic countries with less structured laboratory systems.

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Section: Discussionmentioning

confidence: 99%

Field-Deployable Reverse Transcription-Insulated Isothermal PCR (RT-iiPCR) Assay for Rapid and Sensitive Detection of Foot-and-Mouth Disease Virus

Ambagala

Fisher

Goolia

et al. 2016

Transbound Emerg Dis

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“…A cartridge-based portable RT-PCR assay using dried reagents has been developed by Idaho Technologies (Salt Lake City, UT). Takekawa et al used this assay for the detection of H5N1 in cloacal specimens collected in the field from wild birds and compared it with culture and a laboratory-based PCR assay 52 . The portable assay had equivalent specificity to virus isolation with no false positives, but the sensitivity was lower for specimens with low viral loads.…”

Section: Influenza Virusmentioning

confidence: 99%

Molecular diagnosis of respiratory virus infections

Mahony¹,

Petrich²,

Śmieja³

2011

Critical Reviews in Clinical Laboratory Sciences

193

181

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The appearance of eight new respiratory viruses, including the SARS coronavirus in 2003 and swine-origin influenza A/H1N1 in 2009, in the human population in the past nine years has tested the ability of virology laboratories to develop diagnostic tests to identify these viruses. Nucleic acid based amplification tests (NATs) for respiratory viruses were first introduced two decades ago and today are utilized for the detection of both conventional and emerging viruses. These tests are more sensitive than other diagnostic approaches, including virus isolation in cell culture, shell vial culture (SVC), antigen detection by direct fluorescent antibody (DFA) staining, and rapid enzyme immunoassay (EIA), and now form the backbone of clinical virology laboratory testing around the world. NATs not only provide fast, accurate and sensitive detection of respiratory viruses in clinical specimens but also have increased our understanding of the epidemiology of both new emerging viruses such as the pandemic H1N1 influenza virus of 2009, and conventional viruses such as the common cold viruses, including rhinovirus and coronavirus. Multiplex polymerase chain reaction (PCR) assays introduced in the last five years detect up to 19 different viruses in a single test. Several multiplex PCR tests are now commercially available and tests are working their way into clinical laboratories. The final chapter in the evolution of respiratory virus diagnostics has been the addition of allelic discrimination and detection of single nucleotide polymorphisms associated with antiviral resistance. These assays are now being multiplexed with primary detection and subtyping assays, especially in the case of influenza virus. These resistance assays, together with viral load assays, will enable clinical laboratories to provide physicians with new and important information for optimal treatment of respiratory virus infections.

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“…There are other platforms that do not include nucleic acid extraction step (need to be done separately), such as genesig (Primerdesign Ltd, United Kingdom), Genedrive (Epistem Ltd, Manchester, United Kingdom), Cepheid SmartCycler (Cepheid), T-COR 8 (Tetracore), and R.A.P.I.D. (Idaho Technologies) (Takekawa et al, 2010(Takekawa et al, ,2011. The genesig is now supplying lyophilized qPCR assay kits for 62 bovine, 42 equine, 47 porcine, 60 avian, 40 canine, and 26 feline different pathogens.…”

Section: Point-of-care Diagnosticsmentioning

confidence: 99%