Comparison of real-time RT-PCR, shell vial culture, and conventional cell culture for the detection of the pandemic influenza A (H1N1) in hospitalized patients

Roa, Paula López; Catalán, Pilar; Giannella, Maddalena; Viedma, Darío García de; Sandonís, Virginia; Bouza, Emilio

doi:10.1016/j.diagmicrobio.2010.11.007

Cited by 32 publications

(15 citation statements)

References 14 publications

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“…The major cell types that have been used are Madin Darby canine kidney (MDCK), monkey kidney cells and A549 cells. Virus samples are used to infect the established cell lines for 7-10 days to see the cytopathic effect on cells and different cells show different levels of effect with 100% specificity and 86-94% sensitivity [33]. New commercial mixed cell lines (R-Mix cells, R-mix Too) are also available, which have more sensitivity than other cell lines and take less time.…”

Section: Virus Culturementioning

confidence: 99%

Detection methods for influenza A H1N1 virus with special reference to biosensors: a review

et al. 2020

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H1N1 (Swine flu) is caused by influenza A virus, which is a member of Orthomyxoviridae family. Transmission of H1N1 occurs from human to human through air or sometimes from pigs to humans. The influenza virus has different RNA segments, which can reassert to make new virus strain with the possibility to create an outbreak in unimmunized people. Gene reassortment is a process through which new strains are emerging in pigs, as it has specific receptors for both human influenza and avian influenza viruses. H1N1 binds specifically with an α-2,6 glycosidic bond, which is present in human respiratory tract cells as well as in pigs. Considering the fact of fast multiplication of viruses inside the living cells, rapid detection methods need an hour. Currently, WHO recommended methods for the detection of swine flu include real-time PCR in specific testing centres that take 3–4 h. More recently, a number of methods such as Antigen–Antibody or RT-LAMP and DNA biosensors have also been developed that are rapid and more sensitive. This review describes the various challenges in the diagnosis of H1N1, and merits and demerits of conventional vis-à-vis latest methods with special emphasis on biosensors.

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Section: Virus Culturementioning

confidence: 99%

Detection methods for influenza A H1N1 virus with special reference to biosensors: a review

et al. 2020

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“…9,13 The advantages of real-time RT-PCR over other diagnostic methods include its high sensitivity, high specificity, and rapid turnaround time. 3,10,12,16 To assess various diagnostic tests used for detecting CIV in dogs, the present study sought to 1) estimate and compare the sensitivities of CIV testing methods (VI, FluAB, and real-time RT-PCR) and 2) evaluate the performance of the Flu Detect RIDT in detecting CIV nasal shedding in high-risk shelter dogs.…”

mentioning

confidence: 99%

Evaluation of virus isolation, one-step real-time reverse transcription polymerase chain reaction assay, and two rapid influenza diagnostic tests for detecting canine Influenza A virus H3N8 shedding in dogs

et al. 2013

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Sustained transmission of canine Influenza A virus (CIV) H3N8 among U.S. dogs underscores the threat influenza continues to pose to canine health. Because rapid and accurate detection of infection is critical to the diagnosis and control of CIV, the 2 main objectives of the current study were to estimate and compare the sensitivities of CIV testing methods on canine swab samples and to evaluate the performance of Flu Detect™ (Synbiotics Corp., Kansas City, MO) for detecting CIV nasal shedding in high-risk shelter dogs. To address the first objective, nasal and pharyngeal swab samples were collected from 124 shelter and household dogs seen by Colorado State University Veterinary Teaching Hospital clinicians for canine infectious respiratory disease between April 2006 and March 2007 and tested for CIV shedding using virus isolation, the rapid influenza diagnostic test Directigen Flu A+B™ (BD Diagnostic Systems, Sparks, MD), and real-time reverse transcription polymerase chain reaction (RT-PCR). For the second objective, 1,372 dogs with unknown respiratory health status were sampled from 6 U.S. shelters from December 2009 to November 2010. Samples were tested for presence of CIV using real-time RT-PCR and Flu Detect. Using a stochastic latent class modeling approach, the median sensitivities of virus isolation, rapid influenza diagnostic test, and real-time RT-PCR were 72%, 65%, and 95%, respectively. The Flu Detect test performed poorly for detecting CIV nasal shedding compared to real-time RT-PCR. In conclusion, the real-time RT-PCR has the highest sensitivity for detecting virus nasal shedding and can be used as a rapid diagnostic test for CIV.

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“…Although RT-PCR has excellent test characteristics compared with other diagnostic systems [141,142], it is unclear how different test algorithms actually impact patient care. Some work has been done on the economic impact of influenza RT-PCR testing and publications cite turn-around times as measurable statistics, and although some preliminary work suggests a patient benefit to early diagnosis and antiviral therapy [143,144], the impact of utilizing RT-PCR for influenza A to guide patient care and antibiotic/antiviral utilization has not been completely elucidated [145,146].…”

Section: Practical Issues For Implementation Of Nats In Molecular Diamentioning

confidence: 99%

Rethinking Approaches to Improve the Utilization of Nucleic Acid Amplification Tests for Detection and Characterization of Influenza A in Diagnostic and Reference Laboratories

Wong

Drews²

2011

Future Microbiol.

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Influenza A virus (IFVA) is a significant cause of respiratory infections worldwide and was also responsible for a recent pandemic in 2009. Laboratory identification of IFVA can guide antiviral therapy, assist in cohorting of patients and prevent antibiotic use. Characterization of the virus can track the emergence of novel strains, identify resistance and determine how circulating strains match with vaccine components. The gold standard for detection and characterization of IFVA is nucleic acid amplification technology (e.g., reverse transcriptase PCR [RT-PCR]), which must contend with a constantly evolving viral genome. Although molecular technology has been available for over two decades, there is still an operational gap between assay design and utilization of these tests for the diagnosis and characterization of IFVA. This review will discuss issues surrounding the implementation and use of RT-PCR for the identification and characterization of IFVA, and speculate on why RT-PCR has not been used more widely in clinical laboratories or moved closer to the patient. Newer, less widely used technologies that may change our laboratory practices will be identified and the authors will close with an attempt to identify some future applications of RT-PCR-based technologies for the detection and characterization of IFVA.

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Comparison of real-time RT-PCR, shell vial culture, and conventional cell culture for the detection of the pandemic influenza A (H1N1) in hospitalized patients

Cited by 32 publications

References 14 publications

Detection methods for influenza A H1N1 virus with special reference to biosensors: a review

Detection methods for influenza A H1N1 virus with special reference to biosensors: a review

Evaluation of virus isolation, one-step real-time reverse transcription polymerase chain reaction assay, and two rapid influenza diagnostic tests for detecting canine Influenza A virus H3N8 shedding in dogs

Rethinking Approaches to Improve the Utilization of Nucleic Acid Amplification Tests for Detection and Characterization of Influenza A in Diagnostic and Reference Laboratories

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