Abstract
Duck Tembusu virus (DTMUV) is an emerging pathogenic flavivirus that has resulted in large economic losses to the duck-rearing industry in China since 2010. Therefore, an effective diagnostic approach to monitor the spread of DTMUV is necessary. Here, a novel diagnostic immunochromatographic strip (ICS) assay was developed to detect DTMUV. The assay was carried out using colloidal gold coated with purified monoclonal antibody A12D3 against envelope E protein. Purified polyclonal C12D1 antibodies from BALB/c mice against the envelope E protein were used as the capture antibody. Goat anti-mouse IgG was used to detect DTMUV, which was also assembled on the ICS. Results showed that the ICS could specifically detect DTMUV within 10 min. It also could be stored 25 and 4°C for 4 and 6 months, respectively. The sensitivity of the ICS indicated that the dilution multiples of positive allantoic fluid of DTMUV (LD50: 104.33/0.2 ml) was up to 200. Its specificity and sensibility showed no significant change under the above storage situations. Fifty clinical samples were simultaneously detected by ICS and reverse-transcription polymerase chain reaction with a 93.9% coincidence rate between them. It proved that the ICS in the present study was highly specific, sensitive, repeatable, and more convenient to rapidly detect DTMUV in clinical samples.
Highlights
Like other flaviviruses, Duck Tembusu virus (DTMUV) from the Flaviviridae family has a 10,990nt positive-sense single-stranded RNA genome and belongs to the Ntaya antigenic group
Avian reovirus virus (ARV), H9N2 avian influenza virus (AIV), DTMUV, duck plague virus (DPV), Duck hepatitis A virus type-1 (DHAV-1), egg drop syndrome virus (EDSV), Muscovy duck parovirus (MPV), and Newcastle disease virus (NDV) reference strains were obtained from our laboratory, the Poultry Disease Lab of Shandong Agricultural University, and used to test the specificity of immunochromatographic strip (ICS)
The results showed that the ICS only reacted to the positive allantoic fluid samples of DTMUV, and had no cross reaction with NDV, assay at 4 and 25◦C for 1–10 months
Summary
Duck Tembusu virus (DTMUV) from the Flaviviridae family has a 10,990nt positive-sense single-stranded RNA genome and belongs to the Ntaya antigenic group. In China, it was originally detected in Shanghai in April 2010 and spread rapidly throughout most duck-producing provinces such as Guangdong, Fujian, Jiangsu, Anhui, Henan, and Shandong (Su et al, 2011; Tang et al, 2013). This virus can cause retarded growth, high fever, loss of appetite, severe duck-drop syndrome, and death of the ducks (Yan et al, 2011). Over 10 million ducks have been infected, and Tembusu Virus With ICS more than 1 million ducks died in 2010, which caused massive economic losses in duck-producing industries (Li et al, 2012; He et al, 2017). A rapid detection method is necessary to monitor viral occurrence and prevalence
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