Limits...
New oligonucleotide microarray for rapid diagnosis of avian viral diseases

View Article: PubMed Central - PubMed

ABSTRACT

Background: We developed a new oligonucleotide microarray comprising 16 identical subarrays for simultaneous rapid detection of avian viruses: avian influenza virus (AIV), Newcastle disease virus (NDV), infection bronchitis virus (IBV), and infectious bursal disease virus (IBDV) in single- and mixed-virus infections. The objective of the study was to develop an oligonucleotide microarray for rapid diagnosis of avian diseases that would be used in the course of mass analysis for routine epidemiological surveillance owing to its ability to test one specimen for several infections.

Methods and results: The paper describes the technique for rapid and simultaneous diagnosis of avian diseases such as avian influenza, Newcastle disease, infectious bronchitis and infectious bursal disease with use of oligonucleotide microarray, conditions for hybridization of fluorescent-labelled viral cDNA on the microarray and its specificity tested with use of AIV, NDV, IBV, IBDV strains as well as biomaterials from poultry.

Methods and results: Sensitivity and specificity of the developed microarray was evaluated with use of 122 specimens of biological material: 44 cloacal swabs from sick birds and 78 tissue specimens from dead wild and domestic birds, as well as with use of 15 AIV, NDV, IBV and IBDV strains, different in their origin, epidemiological and biological characteristics (RIBSP Microbial Collection). This microarray demonstrates high diagnostic sensitivity (99.16% within 95% CI limits 97.36–100%) and specificity (100%). Specificity of the developed technique was confirmed by direct sequencing of NP and M (AIV), VP2 (IBDV), S1 (IBV), NP (NDV) gene fragments.

Conclusion: Diagnostic effectiveness of the developed DNA microarray is 99.18% and therefore it can be used in mass survey for specific detection of AIV, NDV, IBV and IBDV circulating in the region in the course of epidemiological surveillance. Rather simple method for rapid diagnosis of avian viral diseases that several times shortens duration of assay versus classical diagnostic methods is proposed.

No MeSH data available.


Intensity profiles of hybridization signals of labeled cDNAs of strains "A/duck/Alberta/35/76" (H1N1) (AIV); "A/duck/Germany/215" (H2N3) (AIV); "A/duck/California/72" (H3N8) (AIV); "A/duck/Czechoslovakia/56" (H4N6) (AIV); "Vinterfild"(IBDV); "BG" (IBDV); "201" (IBDV); "Koktal" (IBDV); "52-70" (IBDV); "H-120" (IBV); "10-95" (IBV); "63/00" (NDV); "52/98" (NDV); "Bor-74 VGNKI" (NDV); "Columba livia /KZ/EKO/15/2014"(NDV)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC5382490&req=5

Fig4: Intensity profiles of hybridization signals of labeled cDNAs of strains "A/duck/Alberta/35/76" (H1N1) (AIV); "A/duck/Germany/215" (H2N3) (AIV); "A/duck/California/72" (H3N8) (AIV); "A/duck/Czechoslovakia/56" (H4N6) (AIV); "Vinterfild"(IBDV); "BG" (IBDV); "201" (IBDV); "Koktal" (IBDV); "52-70" (IBDV); "H-120" (IBV); "10-95" (IBV); "63/00" (NDV); "52/98" (NDV); "Bor-74 VGNKI" (NDV); "Columba livia /KZ/EKO/15/2014"(NDV)

Mentions: The DNA-chip was scanned with InnoScan710AL (“Innopsys”, France) by Cy5 channel activation. The results were interpreted with use of Mapix ver. 5.5.0 software. The signal exceeding the background value was adopted as a positive result. The finding of the study considered reliable if in the course of scanning by Cy5 channel bright fluorescent spots were observed. In the assay of samples the value of specific fluorescence reliably exceeded the value of the background signal (P < 0.05) (Figs. 3 and 4).Fig. 3


New oligonucleotide microarray for rapid diagnosis of avian viral diseases
Intensity profiles of hybridization signals of labeled cDNAs of strains "A/duck/Alberta/35/76" (H1N1) (AIV); "A/duck/Germany/215" (H2N3) (AIV); "A/duck/California/72" (H3N8) (AIV); "A/duck/Czechoslovakia/56" (H4N6) (AIV); "Vinterfild"(IBDV); "BG" (IBDV); "201" (IBDV); "Koktal" (IBDV); "52-70" (IBDV); "H-120" (IBV); "10-95" (IBV); "63/00" (NDV); "52/98" (NDV); "Bor-74 VGNKI" (NDV); "Columba livia /KZ/EKO/15/2014"(NDV)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5382490&req=5

Fig4: Intensity profiles of hybridization signals of labeled cDNAs of strains "A/duck/Alberta/35/76" (H1N1) (AIV); "A/duck/Germany/215" (H2N3) (AIV); "A/duck/California/72" (H3N8) (AIV); "A/duck/Czechoslovakia/56" (H4N6) (AIV); "Vinterfild"(IBDV); "BG" (IBDV); "201" (IBDV); "Koktal" (IBDV); "52-70" (IBDV); "H-120" (IBV); "10-95" (IBV); "63/00" (NDV); "52/98" (NDV); "Bor-74 VGNKI" (NDV); "Columba livia /KZ/EKO/15/2014"(NDV)
Mentions: The DNA-chip was scanned with InnoScan710AL (“Innopsys”, France) by Cy5 channel activation. The results were interpreted with use of Mapix ver. 5.5.0 software. The signal exceeding the background value was adopted as a positive result. The finding of the study considered reliable if in the course of scanning by Cy5 channel bright fluorescent spots were observed. In the assay of samples the value of specific fluorescence reliably exceeded the value of the background signal (P < 0.05) (Figs. 3 and 4).Fig. 3

View Article: PubMed Central - PubMed

ABSTRACT

Background: We developed a new oligonucleotide microarray comprising 16 identical subarrays for simultaneous rapid detection of avian viruses: avian influenza virus (AIV), Newcastle disease virus (NDV), infection bronchitis virus (IBV), and infectious bursal disease virus (IBDV) in single- and mixed-virus infections. The objective of the study was to develop an oligonucleotide microarray for rapid diagnosis of avian diseases that would be used in the course of mass analysis for routine epidemiological surveillance owing to its ability to test one specimen for several infections.

Methods and results: The paper describes the technique for rapid and simultaneous diagnosis of avian diseases such as avian influenza, Newcastle disease, infectious bronchitis and infectious bursal disease with use of oligonucleotide microarray, conditions for hybridization of fluorescent-labelled viral cDNA on the microarray and its specificity tested with use of AIV, NDV, IBV, IBDV strains as well as biomaterials from poultry.

Methods and results: Sensitivity and specificity of the developed microarray was evaluated with use of 122 specimens of biological material: 44 cloacal swabs from sick birds and 78 tissue specimens from dead wild and domestic birds, as well as with use of 15 AIV, NDV, IBV and IBDV strains, different in their origin, epidemiological and biological characteristics (RIBSP Microbial Collection). This microarray demonstrates high diagnostic sensitivity (99.16% within 95% CI limits 97.36&ndash;100%) and specificity (100%). Specificity of the developed technique was confirmed by direct sequencing of NP and M (AIV), VP2 (IBDV), S1 (IBV), NP (NDV) gene fragments.

Conclusion: Diagnostic effectiveness of the developed DNA microarray is 99.18% and therefore it can be used in mass survey for specific detection of AIV, NDV, IBV and IBDV circulating in the region in the course of epidemiological surveillance. Rather simple method for rapid diagnosis of avian viral diseases that several times shortens duration of assay versus classical diagnostic methods is proposed.

No MeSH data available.