Development and evaluation of a gold nanoparticle-based immunochromatographic strip test for the detection of canine parvovirus

Abstract

Canine parvovirus (CPV) is the leading viral cause of enteritis in dogs and occurs mainly in 6- to 8-week-old pups. Rapid diagnosis of CPV under field conditions is now possible due to commercially available immunochromatographic (IC) assays. However, these commercial kits are somewhat expensive because they utilize a minimum of two monoclonal antibodies (mAbs) targeting different epitopes as capture and detector antibodies. Using only a single mAb for both capture and detection purpose may reduce the sensitivity of the assay. In the present study, efforts were made to develop an economical assay that can be utilized for diagnosis of CPV under Indian conditions with a high level of confidence. Rabbit polyclonal antibodies (pAbs) generated against recombinant truncated VP2 proteins of CPV were used as capture antibodies because they can be produced economically, while a commercial anti-CPV mAb was used as the detector antibody. The detection limit of the test strip was 6.6×10 TCID/ml, and it specifically detected CPV-2, CPV-2a and CPV-2b while displaying no cross-reactivity with other common canine enteric pathogens. The relative sensitivity/specificity of pAb based strip test was 71%/92% and 71%/100% in relation to the hemagglutination test and a commercial IC kit, respectively, with substantial agreement. In addition, two commercially available mAbs targeting different epitopes were also used for development of another IC assay, which showed sensitivity, and specificity of 82%/87% and 90%/98% in relation to the hemagglutination test and commercial kit. Hence, the present strip test based on a combination of mAb and pAb provides an acceptable alternative for onsite and cost-effective diagnosis of CPV infection.

Development of an Immunochromatographic Strip for Rapid Detection of Canine Adenovirus

“…In this study, we chose two mAbs which targeted different epitopes as capture and detector antibodies in order to guarantee the sensitivity and specificity of the ICS and prevent other possible antigens from conjugating to the antibody. The limit of detection of the test strip was 2.0 × 10 2 TCID 50 /ml, which is more sensitive than the ICS developed using a single polyclonal antibody (pAb) and single mAb (limit of detection = 6.6 × 10 5 TCID 50 /ml) (Sharma et al, 2018) or another using two mAbs (limit of detection = 2.9 × 10 3 TCID 50 /ml) (Li et al, 2011). Its sensitivity was also higher than that of the commercial CAdV-2 test strip from Korean GenBody (limit of detection = 3.0 × 10 2 TCID 50 /ml).…”

“…Virus isolation and real-time PCR assays are reliable methods for diagnosis of CAdV infections; however, these methods are mostly restricted to well-equipped laboratories. The ICS method has been widely applied to detect various infectious agents (Li et al, 2011;Nakayama et al, 2014;Liu et al, 2019), including canine viruses such as rabies and CPV (Tao and Li, 2014;Sharma et al, 2018). However, there are no publications regarding ICS detection of CAdV, although there is commercial ICS that detect CAdV-2 on the market.…”

Assessing HeberFast® Line Gavac, a lateral flow immunochromatographic system for the rapid detection of anti-Bm86 antibodies in Gavac vaccinated cattle

“…In veterinary medicine, lateral flow immunoassays have been developed for various purposes; detection of diseases in companion and farm animals to achieve better results in the care and development of animals, for example, Parvovirus 21 , Newcastle 22 , Avian infectious bronchitis virus 23 , African swine fever 24 and Babesiosis 25 , The rapid detection of these diseases reduces the economic losses. Immunochromatographic methods have also been developed to detect antibiotics and analogs in milk 26 , which is greatly important for food safety.…”

One-Step Rapid Quantification of Serum Neutralizing Antibody after COVID-19 Vaccination by a High-Throughput Nanoplasmonic Sensor Platform

“…27,[35][36] GNPs in the range of 20-40 nm in diameter were generally recommended for use in biological detection procedures due to their sufficient stability and immune reactivity. [37][38][39] However, only large GNPs that fit in the nanocup size can effectively help to increase the intensity of the localized electric field and improve the detection sensitivity. [40][41] Thus, NHGNPs with higher stability and larger diameter (~100 nm) will be a potential strategy to improve the SPR effect on the nanocup sensor chip.…”