Open Access
Research (Published online: 02-08-2017)
2. Preparation and evaluation of Salmonella Enteritidis antigen conjugated with nanogold for screening of poultry flocks
Hazem Mohammed Ibrahim, Rafik Hamed Sayed, Wafaa Ragab Abdel-Aziz and Rafik Tawfik Soliman
Veterinary World, 10(8): 848-853

Hazem Mohammed Ibrahim: Department of Bacterial Sera and Antigen Research, Veterinary Serum and Vaccine Research Institute, Abbasia, Cairo, Egypt.
Rafik Hamed Sayed: Central Laboratory for Evaluation of Veterinary Biologics, Abbasia, Cairo, Egypt.
Wafaa Ragab Abdel-Aziz: Department of Bacterial Sera and Antigen Research, Veterinary Serum and Vaccine Research Institute, Abbasia, Cairo, Egypt.
Rafik Tawfik Soliman: Department of Microbiology, Faculty of Veterinary Medicine, Cairo University, Cairo, Egypt.

doi: 10.14202/vetworld.2017.848-853

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Article history: Received: 10-04-2017, Accepted: 22-06-2017, Published online: 02-08-2017

Corresponding author: Hazem Mohammed Ibrahim


Citation: Ibrahim HM, Sayed RH, Abdel-Aziz WR, Soliman RT (2017) Preparation and evaluation of Salmonella Enteritidis antigen conjugated with nanogold for screening of poultry flocks, Veterinary World, 10(8): 848-853.

Aim: The present work aimed to develop lateral flow immunochromatographic test (LFIT) for detection of Salmonella Enteritidis (SE) specific antibodies in chicken sera.

Materials and Methods: A rapid LFIT has been developed, in which SE Group D antigen labeled with the gold chloride molecules laid on the conjugate pad. Staphylococcus aureus protein A was used as capture antibody at the test line (T) of a nitrocellulose (NC) membrane and anti-SE antigen-specific rabbit antibodies were used as capture antibody at the control line (C) of the NC strip in the lateral flow layout device.

Results: Using the developed LFIT, the minimal amount of SE-specific antibodies that can be detected in chicken serum sample was 1427 enzyme-linked immunosorbent assay (ELISA) unit/100 μl that was equal to 0.1 μg (Ab)/100 μl sample. 100 suspected serum samples collected from a poultry flock were tested with the prepared SE-LFIT kits and the locally prepared stained Salmonella antigen, and the results were compared with those obtained from examination of these samples with Salmonella Group D antibody ELISA kit as the gold standard test. The sensitivity, specificity, and accuracy of the prepared SE-LFIT antigen kits were 94.4%, 90%, and 94%, respectively, while those obtained with stained Salmonella antigen were 88.8%, 90%, and 89%, respectively.

Conclusion: The developed test is a simple field rapid test of high sensitivity, specificity, and accuracy that can improve and facilitates rapid field surveillance of salmonellosis among chickens.

Keywords: diagnosis, lateral flow, poultry, Salmonella Enteritidis.


1. Borges, K.A., Furian, T.Q., de Souza, S.N., Tondo, E.C., Streck, A.F., Salle, C.T., de Souza Moraes, H.L. and do Nascimento, V.P. (2017) Spread of a major clone of Salmonella Enterica serotype enteritidis in poultry and in Salmonellosis outbreaks in southern Brazil. J. Food Protect., 80(1): 158-163. [Crossref] [PubMed]

2. Jarquin, R., Hanning, I., Ahn, S. and Ricke, S.C. (2009) Development of rapid detection and genetic characterization of Salmonella in poultry breeder feeds. Sens. (Basel), 9(7): 5308-5323. [Crossref] [PubMed] [PMC]

3. Galis, A.M., Marcq, C., Marlier, D., Portetelle, D., Van, I., Beckers, Y. and Thewis, A. (2013) Control of Salmonella contamination of shell eggs-preharvest and postharvest methods: A review. Compr. Rev. Food Sci. Food Saf., 12: 155-182. [Crossref]

4. Wu, B., Zhang, X., Pan, W., Zhang, L. and Zhang, F. (2015) Determination of Salmonella Pullorum with nanoparticles immune based lateral flow strip assay. Adv. Microbiol., 5: 364-369. Available from: Last accessed on 1/11/2016.

5. Chen, W., Rahman, R.T., Antora, R.A., Saqib, N., Hossain, P., Zhang, J., Al-Hajj, N.Q., Lou, Z. and Zaixiang, L. (2015) Lateral flow test strip approaches for rapid detection of food contaminants and pathogens: Review. Am. Res. Thoughts, 1(12): 1222-1248.

6. Lata, K., Naik, L., Sharma, R. and Rajput, Y.S. (2013) Lateral flow assay-concept and its applications in food analysis. Indian Food Ind. Mag., 32(5): 22-32.

7. Pandey, S.K., Suri, C.R., Chaudhry, M., Tiwari, R.P. and Praveen, R. (2012) A gold nanoparticles based immuno-bioprobe for detection of Vi capsular polysaccharide of Salmonella Enterica Serovars typhi. Mol. Biosys., 8: 1853-1860. [Crossref] [PubMed]

8. Shyu, R., Tang, S., Chiao, D. and Hung, Y. (2010) Gold nanoparticle-based lateral flow assay for detection of staphylococcal enterotoxin B. Food Chem., 118: 462-466. [Crossref]

9. Hong, W., Lihua, H., Haoran, W., Jianfeng, Q., Zhaobiao, G., Chengke, X., Ziwen, Z., Youbao, Z., Zongmin, D., Yanfeng, Y., Yan, Z., Huijie, H., Ruifu, Y. and Lei, Z. (2010) Development of an up converting phosphor technology-based 10-channel lateral flow assay for profiling antibodies against Yersinia pestis. J. Microbiol. Methods, 83: 133-140. [Crossref]

10. Chen, W., Zhang, J., Lu, G., Yuan, Z., Wu, Q. and Li, J. (2014) Development of an immunochromatographic lateral flow device for rapid diagnosis of Vibrio cholerae O1 serotype Ogawa. Clin. Biochem., 47: 448-454. [Crossref] [PubMed]

11. Oem, J.K., Ferris, N.P., Kwang-Nyeong, L., Yi-Seok, J., Bang-Hun, H. and Jong-Hyeon, P. (2009) Simple and rapid lateral-flow assay for the detection of foot-and-mouth disease virus. Clin. Vaccine Immunol., 16(11): 1660-1664. [Crossref] [PubMed] [PMC]

12. Shiota, S., Mannen, K. and Matsumoto, T. (2009) Development and evaluation of a rapid neutralizing antibody test for rabies. J. Virol. Methods, 161: 58-62. [Crossref] [PubMed]

13. Yang, W., Li, X., Liu, G., Zhang, B., Zhang, Y., Kong, T., Tang, J., Li, D. and Wang, Z. (2011) A colloidal gold prob-based silver enhancement immune-chromatographic assay for the rapid detection of abrin-a. Biosens. Bioelectron., 26: 3710-3713. [Crossref] [PubMed]

14. Wary, C. and Wary, A. (2000) Salmonella in domestic animals. CABI Publishing, Oxfordshire.

15. Gulbenkian, S., Wharton, J. and Polak, J.M. (1987) The visualization of cardiovascular innervations in the guinea-pig using an antiserum to protein gene product 9.5. J. Auton. Nerv. Syst., 18: 235-247. [Crossref]

16. Donovan, J. and Brown, P. (1995) Current Protocols in Immunology. Wiley Online Library, John Wiley & Sons, Inc., US.

17. Singh, J., Sharma, S. and Seema, N. (2015) Evaluation of gold nanoparticle based lateral flow assays for diagnosis of Enterobacteriaceae members in food and water. Food Chem., 170: 470-483. [Crossref] [PubMed]

18. Singh, J., Sharma, S. and Seema, N. (2015) Nanogold based lateral flow assay for the detection of Salmonella Typhi in environmental water samples. Anal. Methods, 7: 9281-9288. [Crossref]

19. Statistix. (1996) Analytical Software, Version 1.0, Tallahassee, FL, USA.

20. Timmreck, T.C. (1994) An Introduction to Epidemiology. Jones and Bartlett Publishers, Boston, London and Singapore.

21. Thrusfield, M. (2007) Veterinary Epidemiology. 3rd ed. Blackwell Science, London. [PubMed]

22. Biagini, R.E., Sammons, D.L., Smith, J.P., MacKenzie, B.A., Striley, C.A.F., Snawder, J.E., Robertson, S.A. and Quinn, C.P. (2006) Rapid, sensitive and specific lateral flow immunochromatographic device to measure anti-anthrax protective antigen immunoglobulin in serum and whole blood. Clin. Vaccine Immunol., 13(5): 541-546. [Crossref] [PubMed] [PMC]