Point-of-Care and Label-Free Detection of Porcine Reproductive and Respiratory Syndrome and Swine Influenza Viruses Using a Microfluidic Device with Photonic Integrated Circuits
Georgios ManessisMaciej FrantGrzegorz WoźniakowskiLapo NannucciMartina BenedettiLilla DénesGyula BalkaAthanasios Ι. GelasakisC. SquiresS. RecueroCarlos SánchezAmadeu GriolAlessandro GiustiIoannis Bossis
2
Citation
56
Reference
10
Related Paper
Citation Trend
Abstract:
Swine viral diseases challenge the sector’s sustainability by affecting productivity and the health and welfare of the animals. The lack of antiviral drugs and/or effective vaccines renders early and reliable diagnosis the basis of viral disease management, underlining the importance of point-of-care (POC) diagnostics. A novel POC diagnostic device utilizing photonic integrated circuits (PICs), microfluidics, and information and communication technologies for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) and swine influenza A (SIV) was validated using spiked and clinical oral fluid samples. Metrics including sensitivity, specificity, accuracy, precision, positive likelihood ratio (PLR), negative likelihood ratio (NLR), and diagnostic odds ratio (DOR) were calculated to assess the performance of the device. For PRRSV, the device achieved a sensitivity of 83.5%, specificity of 77.8%, and DOR values of 17.66, whereas the values for SIV were 81.8%, 82.2%, and 20.81, respectively. The POC device and PICs can be used for the detection of PRRSV and SIV in the field, paving the way for the introduction of novel technologies in the field of animal POC diagnostics to further optimize livestock biosecurity.Arterivirus
Circovirus
Cite
Citations (12)
A porcine reproductive and respiratory syndrome virus(PRRSV) strain isolated from visceral organs was designated as PRRSV-FS.The sequence analysis revealed that the isolated virus was a mutation strain as the Nsp2 gene existed amino acid deletion of 30 and the GP5 gene was conservative relatively.The genome of PRRSV FS strain showed 88.7%,98.8%,58.5% of nucleotide indentity with ATCC VR-22332,JN-HS,Europena Lelystad virus respectively.
Strain (injury)
Arterivirus
Cite
Citations (0)
An one-step Multiplex RT-PCR assay was developed using two pairs of primers(PH1/PH2 and PT1/PT2) designed according to the conservative sequences of porcine circovirus type 2(PCV2) ORF2 and porcine respiratory and reproductive syndrome virus(PRRSV) ORF5 respectively.PCV2 DNA and PRRSV RNA was extracted from the mixed sample of PCV2 and PRRSV and used as PCR template.Two specific PCR fragments of PCV2 560 bp and PRRSV 398 bp were amplified.No PCR products were detected from other four porcine pathogenic viruses.The assay has a sensitivity of detecting 10 ng of PCV2 DNA and 5 ng PRRSV RNA.
Multiplex
Circovirus
Cite
Citations (0)
In this study,differential centrifugation and discontinuous sucrose density gradient centrifugation were adopted to concentrate and purify porcine reproductive respiratory and syndrome virus(PRRSV),The purified PRRSV which is treated with Trichlor_trifluorethan regard as ELISA_antigen.The indirect ELISA was sensitive and specific,and it was superior to both IPMA and IFA in detecting antibodies in the sensitive.
Differential centrifugation
Cite
Citations (0)
Cite
Citations (34)
A kit for detection of antibody against porcine reproductive and respiratory syndrome virus(PRRSV) was prepared by immunoperoxidase monolayer assay(IPMA) and assembled as PRRSV-IPMA kit.Sensitivity,specificity,repeatability and preservation of the kit were evaluated.Using the PRRSV-IPMA kit sera from PRRSV-infected pigs were detected to be 50% and 100% of positive rates at week 1 and 2 post-inoculation,respectively.The sera from control group pigs showed negative reaction.In comparison,parallel detections were done using the PRRSV-IPMA kit and the commercial PRRSV-ELISA kit from IDEXX Co.,and the agreement rate was 83.3%.The PRRSV-IPMA kit was stable for 18 months at-20℃,and didn't cross-react with the other virus reference sera.PRRSV antibodies in 520 serum samples from 13 farms of Heilongjiang,Jilin,Liaoning,Hebei and Shanghai were detected by the PRRSV-(IPMA) kit.The positive rates of four farms were more than 90%,and only two farms were negative,and 10%-57% in the rest of farms.
Immunoperoxidase
Arterivirus
Cite
Citations (2)
Seroprevalence
Arterivirus
Cite
Citations (8)
Alveolar macrophage
Cite
Citations (10)
A one-step real-time Taqman RT-PCR assay for porcine reproductive and respiratory syndrome virus(PRRSV) was developed to detect the two type of PRRSV,American and European strain,by targeting the ORF7 gene of the virus.This new assay was compared to commercial methods developed by applied biosystems company.Quantitation was performed against a standard samples of the PRRSV for which a minimum of 10 copies per reaction were detected with a corresponding Ct value of 0.74%-1.52% and it shows no cross reaction with other swine pathogens.By detecting 40 clinical samples of the virus,the results are the same as the commercial kits.
TaqMan
Highly pathogenic
Arterivirus
Cite
Citations (0)
Objective: To establish a herd negative for porcine reproductive and respiratory syndrome virus (PRRSV) from PRRSV-positive sources by management of the gilt pool and batching of pig flow. Methods: Two groups of PRRSV-positive gilts were housed in common acclimatization areas for 70 to 100 days, bred in an off-site finishing facility, and farrowed in a separate facility. Piglets weaned at 5 to 7 days of age were moved to off-site nurseries in weekly batches and mixed with cohort sentinel piglets from a PRRSV-negative herd. Each nursery batch was tested for PRRSV twice by PCR and once by ELISA. Two to five negative batches were grouped in off-site grower facilities, and tested again. Negative groups moved to a quarantine facility. Negative quarantine groups were used for stocking the PRRSV-negative herd. Results: Of the 31 batches of nursery pigs produced, three batches, born 2, 4, and 6 weeks after farrowings started, were PRRSV-positive. Six grower groups (comprising 23 nursery batches) that were PRRSV-negative after repeated testing were assembled into five groups in a quarantine facility and remained PRRSV-negative. Two grower groups were rejected. A total of 9500 pigs were produced, from which 3415 pigs were selected to stock the PRRSV-negative herd. Implications: A PRRSV-negative population was established from positive sources by managing the gilt pool and batching the pig flow, allowing for preservation of elite genetics. It appeared that PRRSV infection, indicated by lack of seroconversion in the offspring, eventually either disappeared or became inactive in the donor gilt population.
Cite
Citations (42)