Development of an immunochromatographic strip for the detection of antibodies against foot-and-mouth disease virus serotype O

College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China.
Journal of virological methods (Impact Factor: 1.78). 05/2010; 165(2):139-44. DOI: 10.1016/j.jviromet.2010.01.001
Source: PubMed


An immunochromatographic strip was developed for the serological detection of type O foot-and-mouth disease (FMD) in swine. In the strip, the expressed protein of VP1, the main protective antigen of FMDV, labeled with colloidal gold was used as the detector, the staphylococcal protein A (SPA) and swine anti-foot-and-mouth disease virus (FMDV) antibody were blotted on the nitrocellulose membrane for the test and control lines, respectively. 296 swine serum samples were collected to evaluate the characteristics of the strip in comparison with existing commercial liquid-phage blocking ELISA (LPB ELISA) kit and peptide ELISA kit. The strip was shown to be of high specificity and sensitivity. Furthermore, the dipstick assay based on the strip is rapid (5 min) and easy to perform with no requirement of professional skills, reagents nor equipment. This suggests that the immunochromatographic strip is an acceptable alternative for use in clinical laboratories lacking specialized equipment and for field diagnosis.

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    • "Colloidal gold immunochromatographic assay (GICA) is convenient, is rapid, has high specificity and sensitivity, and can be performed either without instruments or with only a simple instrument, making it suitable for clinical diagnosis and drug testing purposes in almost any context [29-31]. Colloidal gold is a negatively charged hydrophobic rubber particle that maintains a stable colloidal system through electrostatic repulsion [22]. "
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    ABSTRACT: Background The incidence of PHE among pigs in many countries is on the rise, and it has caused great economic losses to the pig industry. Therefore, the development of a sensitive, specific, and easily-performed assay is crucial for the rapid detection and surveillance of PHE-CoV infection and transmission. Results An immunochromatographic strip was developed for the detection of PHE-CoV. The colloidal gold-labeled MAb 4D4 was used as the detection reagent, and the MAb 1E2 and goat anti-mouse IgG coated the strip's test and control lines, respectively. The immunochromatographic strip was capable of specifically detecting PHE-CoV with a HA unit of 2 within 10 min. Storage of the strips at room temperature for 6 months or at 4°C for 12 months did not change their sensitivity or specificity. Using RT-PCR as a reference test, the relative specificity and sensitivity of the immunochromatographic strip were determined to be 100% and 97.78%, respectively. There was an excellent agreement between the results obtained by RT-PCR and the immunochromatographic strips (kappa = 0.976). Additionally, there was a strong agreement between the sandwich enzyme-linked immunosorbent assay (ELISA) and immunochromatographic strips (Kappa = 0.976). When the immunochromatographic strips were used for diagnosing PHE-CoV infection in the Jilin Province, the PHE-CoV-positive rate ranged from 61.54% in the Jilin district to 17.95% in the Songyuan district. Conclusions Based on its high specificity, sensitivity, and stability, the immunochromatographic strip would be suitable for on-site detection of PHE-CoV for surveillance and epidemiological purposes.
    Virology Journal 08/2012; 9(1):172. DOI:10.1186/1743-422X-9-172 · 2.18 Impact Factor
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    • "Colloidal gold was prepared according to the reported method[13]. 2C'3AB protein was diluted serially to give 10,20,30,40,50,60,70 and 80 μg/ml. "
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    ABSTRACT: Foot-and-mouth disease (FMD) is an extremely contagious viral disease of cattle, pigs, sheep, goats, and many cloven-hoofed wild animals. FMDV serotypes O and Asia 1 have circulated separately in China during the last fifty years, and eliminating infected animals and vaccination are the main policies to prevent and control FMD. Antibodies to NSPs exist in infected animals, and were utilized to differentiate between infected and vaccinated animals. The reliability of detection of 3AB or 3ABC antibodies is higher than that of other NSPs. The test of 3AB is still credible because 3C protein's immunogenicity is the weakest. The 2C protein, immediately N-terminal of 3AB, was used to differentiate between infected and vaccinated animals. The use of the immunochromatographic strip is facile for clinical laboratories lacking specialized equipment and for rapid field diagnosis. In this study, an immunochromatographic strip with non-structural protein (NSP) 2C'3AB was developed and validated to differentiate foot-and-mouth disease infected from vaccinated animals. A part of N-terminal of 2C protein gene and whole 3AB gene were connected and prokaryotically expressed as the antigens labeled with colloidal gold was used as the detector, the 2C'3AB protein and rabbits anti-2C'3AB antibodies were blotted on the nitrocellulose(NC) membrane for the test and control lines, respectively. 387 serum samples were collected to evaluate the characteristics of the strip in comparison with existing commercial 3ABC antibody ELISA kit. The coincidence rate of pigs negative serum, pigs vaccinated serum, pigs infected serum was 100%, 97.2%, 95.0%, respectively. The coincidence rate of cattle negative serum, cattle vaccinated serum, cattle infected serum was 100%, 96.7%, 98.0%, respectively. The coincidence rate of sheep negative serum, sheep infected serum was 97.6%, 96.3%, respectively. The strip was shown to be of high specificity and sensitivity, good repeatability and stability. These data suggest that the immunochromatographic strip is a useful tool for rapid on-site diagnosing animals infected foot-and-mouth disease virus.
    Virology Journal 04/2011; 8(1):186. DOI:10.1186/1743-422X-8-186 · 2.18 Impact Factor
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    ABSTRACT: A pocket device for rapid combined test of biomedical samples is comprised of four parts: strip, strip attached column, sampling head and test tube. One to multiple strips are separately attached to the shallow groove on the surface of the strip attached column. The strip attached column is placed inside the test tube of the device. The sampling head is in front of the test tube. The sampling head has a slim tube type suction nozzle, which is capped with a rubber protection cap. The cavity of the sampling head is front-to-back filled with sample filter material and sample water absorbent material. The sealing cap at the end of the test tube is able to block the waste after testing to prevent leakage. During the sample testing, one removes the rubber protection cap of sampling head suction nozzle and dips the suction nozzle into the test sample. Each strip undergoes once sampling and is detected at the same background. In 3-5 minutes we can concurrently test several indexes of the sample in the transparent areas of test tube with naked eyes. This device has many advantages: small size, low cost, simple, safe and fast operation, and it can effectively prevent detection of cross-interference. Post-test waste is still enclosed in the test tube, which helps to prevent the spread of pathogens and iatrogenic infections, and to dispose waste for environmental protection.
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