Rapid detection of influenza virus H1 by the polymerase chain reaction

Bressoud, Albéric; Whitcomb, Jeannette M.; Pourzand, Charareh; Haller, Otto; Cerutti, Peter

doi:10.1016/0006-291x(90)92040-7

Cited by 31 publications

(20 citation statements)

References 13 publications

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“…These viruses present an important diagnostic problem, and the rapid detection, typing, and subtyping of influenza A and B virus strains are of both clinical and epidemiological value. While antibody-based methods still form the foundations of routine diagnostic work, many reports over the last decade have demonstrated the utility and superiority of PCR-based diagnostic (2,4,14,19,22,26) and retrospective (23) tests. Unfortunately PCR-based methods suffer from a problem in that simply producing DNA isn't sufficient evidence that one has amplified the right product.…”

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confidence: 99%

“…Printed arrays were air dried for a few minutes at 50 to 60°C and then stored overnight at 20 to 37°C over desiccant. The arrays were rehydrated for 4 h in a humid atmosphere, dried briefly at 50°C on a heating block, washed once in 0.2% sodium dodecyl sulfate (SDS) and twice in water (1 min each), and treated with sodium borohydride (1.0 g of NaBH 4 [Sigma] dissolved in 300 ml of phosphate-buffered saline plus 100 ml of ethanol [17]) for 5 min. The DNA was denatured in water (2 min at 95°C) and then washed again (once in 0.2% SDS and once in water [1 min each]).…”

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confidence: 99%

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Typing and Subtyping Influenza Virus Using DNA Microarrays and Multiplex Reverse Transcriptase PCR

Chen

Evans³

2001

J Clin Microbiol

152

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A model DNA microarray has been prepared and shown to facilitate typing and subtyping of human influenza A and B viruses. Reverse transcriptase PCR was used to prepare cDNAs encoding ϳ500-bp influenza virus gene fragments, which were then cloned, sequenced, reamplified, and spotted to form a glass-bound microarray. These target DNAs included multiple fragments of the hemagglutinin, neuraminidase, and matrix protein genes. Cy3-or Cy5-labeled fluorescent probes were then hybridized to these target DNAs, and the arrays were scanned to determine the probe binding site(s). The hybridization pattern agreed perfectly with the known grid location of each target, and the signal-to-background ratio varied from 5 to 30. No crosshybridization could be detected beyond that expected from the limited degree of sequence overlap between different probes and targets. At least 100 to 150 bp of homology was required for hybridization under the conditions used in this study. Combinations of Cy3-and Cy5-labeled DNAs can also be hybridized to the same chip, permitting further differentiation of amplified molecules in complex mixtures. In a more realistic test of the technology, several sets of multiplex PCR primers that collectively target influenza A and B virus strains were identified and were used to type and subtype several previously unsequenced influenza virus isolates. The results show that DNA microarray technology provides a useful supplement to PCR-based diagnostic methods.

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Typing and Subtyping Influenza Virus Using DNA Microarrays and Multiplex Reverse Transcriptase PCR

Chen

Evans³

2001

J Clin Microbiol

152

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“…A similar approach has been used to show clearance of an avian influenza virus from ducks (Wang & Webster, 1990), and to demonstrate virus in nasal washings from human patients (Bressoud et al, 1990). The almost exclusive presence of replicating influenza virus in the mouse lung following i.n.…”

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confidence: 99%

Influenza virus RNA in the lung and lymphoid tissue of immunologically intact and CD4-depleted mice

Eichelberger

Wang

Allan

et al. 1991

Journal of General Virology

105

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“…PCR assays have been designed for either Type A only (Pisareva et al, 1992;Cherian et al, 1994), or for distinguishing between Type A, B or C viruses (Claas et al, 1992;Claas et al, 1993) or indeed for the diagnosis of human influenza. Subtype specific primers have been designed to detect single subtypes of Type A influenza viruses (Bressoud et al, 1990) or to differentiate between subtypes of HAs (Lee et al, 2001;Yamada et al, 1991;Zhang et al, 1991) and NAs (Zhang et al, 1991). The study aimed at the establishment of a rapid method for the screening or detection of not only the MP genes, the relatively highly conserved gene in all Type A viruses (Lamb and Krug, 1996) but also the H 5 HA in a single reaction.…”

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confidence: 99%

Multiplex RT-PCR for the rapid detection of matrix protein and H5 genes of highly pathogenic avian influenza virus

2016

Int. J. Curr. Res. Biol. Med.

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Avian influenza (AI) is an economic, zoonotically important disease in worldwide. It is an infectious disease of birds caused by influenza virus which also affect human beings. Outbreaks of H 5 N 1 highly pathogenic avian influenza (HPAI) virus caused great economic losses to the poultry industry. Rapid typing and subtyping of HPAI viruses are desirable for taking prompt control measure to prevent the spread of the disease. Here, we develop multiplex RT-PCR technique for the rapid, simultaneous detection of Matrix protein (MP) and H 5 genes of HPAI from RNA extracted from tracheal swabs using Finnzymes Phusion TM Flash HighFidelity PCR Master Mix (2x) based on modified Phusion Hot Start DNA Polymerase and QIAgen RT-PCR Kit. Reverse transcription and PCR were carried out with a mixture of primers specific for influenza viruses of type A subtype H 5 in a single reaction system under identical conditions. Both the PCR detection systems amplified matrix protein and H 5 genes specific 245bp and 545bp bands, respectively. However, QIAgen RT-PCR Kit required 3 hours to complete the PCR cycle, whereas Phusion Flash method required 1 hour and 15 minutes only. On the other hand, multiplex PCR was finished in one tube for both MP and H 5 genes. Therefore, it is time and cost effective. Phusion Flash method required relatively less time and cost. After validation with few more field samples, it can be used for the rapid, simultaneous detection of MP and H 5 genes of HPAI in the field condition.

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Rapid detection of influenza virus H1 by the polymerase chain reaction

Cited by 31 publications

References 13 publications

Typing and Subtyping Influenza Virus Using DNA Microarrays and Multiplex Reverse Transcriptase PCR

Typing and Subtyping Influenza Virus Using DNA Microarrays and Multiplex Reverse Transcriptase PCR

Influenza virus RNA in the lung and lymphoid tissue of immunologically intact and CD4-depleted mice

Multiplex RT-PCR for the rapid detection of matrix protein and H5 genes of highly pathogenic avian influenza virus

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