Improved detection of human influenza A and B viruses in respiratory tract specimens by hemi-nested PCR

Claudia Minosse, Marina Selleri, Maria S. Zaniratti, Francesco N. Lauria, Vincenzo Puro, Fabrizio Carletti, Giuseppina Cappiello, Gina Gualano, Nazario Bevilacqua, Maria R. Capobianchi

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6 Citations (Scopus)

Abstract

RT-PCR is the most sensitive assay for diagnosis of influenza, due to enhanced rapidity and sensitivity as compared to classical methods. Hemi-nested RT-PCR was developed, targeting NP gene for influenza A and NS gene for influenza B, based on a previous single round RT-PCR method. The new method was compared with the previous technique for analytical sensitivity and specificity, and was applied to clinical samples from the lower and upper respiratory tract. The analytical sensitivity of hemi-nested RT-PCR was 10 (influenza A) and 4 times (influenza B) higher than the previous method. A high specificity of the new hemi-nested RT-PCR assay was observed by using whole respiratory viruses. When applied to lower respiratory tract specimens, the new method showed an increased rate of positivity as compared to the previous technique (9.3% versus 0.7% for influenza A, and 0.9% versus 0.2% for influenza B). Screening of upper respiratory tract samples collected during the seasonal 2005-2006 outbreak indicated 26.4% and 5.8% positivity for influenza A and B, respectively. The results were confirmed by sequence analysis: apart from influenza B, both influenza A subtypes H3N2 and H1N1, associated with the seasonal outbreak, were detected.

Original languageEnglish
Pages (from-to)225-228
Number of pages4
JournalJournal of Virological Methods
Volume141
Issue number2
DOIs
Publication statusPublished - May 2007

Fingerprint

Influenza B virus
Influenza A virus
Respiratory System
Human Influenza
Polymerase Chain Reaction
Disease Outbreaks
Gene Targeting
Sequence Analysis
Viruses

Keywords

  • Influenza
  • Molecular diagnosis
  • Respiratory infections
  • RT-PCR

ASJC Scopus subject areas

  • Virology

Cite this

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title = "Improved detection of human influenza A and B viruses in respiratory tract specimens by hemi-nested PCR",
abstract = "RT-PCR is the most sensitive assay for diagnosis of influenza, due to enhanced rapidity and sensitivity as compared to classical methods. Hemi-nested RT-PCR was developed, targeting NP gene for influenza A and NS gene for influenza B, based on a previous single round RT-PCR method. The new method was compared with the previous technique for analytical sensitivity and specificity, and was applied to clinical samples from the lower and upper respiratory tract. The analytical sensitivity of hemi-nested RT-PCR was 10 (influenza A) and 4 times (influenza B) higher than the previous method. A high specificity of the new hemi-nested RT-PCR assay was observed by using whole respiratory viruses. When applied to lower respiratory tract specimens, the new method showed an increased rate of positivity as compared to the previous technique (9.3{\%} versus 0.7{\%} for influenza A, and 0.9{\%} versus 0.2{\%} for influenza B). Screening of upper respiratory tract samples collected during the seasonal 2005-2006 outbreak indicated 26.4{\%} and 5.8{\%} positivity for influenza A and B, respectively. The results were confirmed by sequence analysis: apart from influenza B, both influenza A subtypes H3N2 and H1N1, associated with the seasonal outbreak, were detected.",
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T1 - Improved detection of human influenza A and B viruses in respiratory tract specimens by hemi-nested PCR

AU - Minosse, Claudia

AU - Selleri, Marina

AU - Zaniratti, Maria S.

AU - Lauria, Francesco N.

AU - Puro, Vincenzo

AU - Carletti, Fabrizio

AU - Cappiello, Giuseppina

AU - Gualano, Gina

AU - Bevilacqua, Nazario

AU - Capobianchi, Maria R.

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AB - RT-PCR is the most sensitive assay for diagnosis of influenza, due to enhanced rapidity and sensitivity as compared to classical methods. Hemi-nested RT-PCR was developed, targeting NP gene for influenza A and NS gene for influenza B, based on a previous single round RT-PCR method. The new method was compared with the previous technique for analytical sensitivity and specificity, and was applied to clinical samples from the lower and upper respiratory tract. The analytical sensitivity of hemi-nested RT-PCR was 10 (influenza A) and 4 times (influenza B) higher than the previous method. A high specificity of the new hemi-nested RT-PCR assay was observed by using whole respiratory viruses. When applied to lower respiratory tract specimens, the new method showed an increased rate of positivity as compared to the previous technique (9.3% versus 0.7% for influenza A, and 0.9% versus 0.2% for influenza B). Screening of upper respiratory tract samples collected during the seasonal 2005-2006 outbreak indicated 26.4% and 5.8% positivity for influenza A and B, respectively. The results were confirmed by sequence analysis: apart from influenza B, both influenza A subtypes H3N2 and H1N1, associated with the seasonal outbreak, were detected.

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