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 language | English |
|---|---|
| Pages (from-to) | 225-228 |
| Number of pages | 4 |
| Journal | Journal of Virological Methods |
| Volume | 141 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - May 2007 |
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Keywords
- Influenza
- Molecular diagnosis
- Respiratory infections
- RT-PCR
ASJC Scopus subject areas
- Virology
Cite this
Improved detection of human influenza A and B viruses in respiratory tract specimens by hemi-nested PCR. / Minosse, Claudia; Selleri, Marina; Zaniratti, Maria S.; Lauria, Francesco N.; Puro, Vincenzo; Carletti, Fabrizio; Cappiello, Giuseppina; Gualano, Gina; Bevilacqua, Nazario; Capobianchi, Maria R.
In: Journal of Virological Methods, Vol. 141, No. 2, 05.2007, p. 225-228.Research output: Contribution to journal › Article
}
TY - JOUR
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.
PY - 2007/5
Y1 - 2007/5
N2 - 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.
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.
KW - Influenza
KW - Molecular diagnosis
KW - Respiratory infections
KW - RT-PCR
UR - http://www.scopus.com/inward/record.url?scp=33947525828&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33947525828&partnerID=8YFLogxK
U2 - 10.1016/j.jviromet.2006.12.010
DO - 10.1016/j.jviromet.2006.12.010
M3 - Article
C2 - 17257688
AN - SCOPUS:33947525828
VL - 141
SP - 225
EP - 228
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 2
ER -