Validation of an enzyme-linked immunosorbent assay for diagnosis of human trichinellosis - PubMed (original) (raw)

Validation of an enzyme-linked immunosorbent assay for diagnosis of human trichinellosis

Maria Angeles Gómez-Morales et al. Clin Vaccine Immunol. 2008 Nov.

Abstract

Trichinellosis is a zoonotic disease caused by the consumption of raw or semiraw meat from different animals harboring Trichinella larvae in their muscles. Since there are no pathognomonic signs, diagnosis can be difficult; for this reason, serology is important. The objective of this study was to validate an enzyme-linked immunosorbent assay (ELISA) using excretory/secretory antigens to detect anti-Trichinella immunoglobulin G antibodies in human sera. A total of 3,505 human serum samples were tested. A receiver-operator characteristic (ROC) curve analysis was performed. The accuracy of the test was determined by calculating the area under the curve, which was equal to 0.999, indicating high accuracy. The coefficient of variation calculated for data from four serum samples in eight working sessions was no higher than 5% for the positive sera or 14% for the negative sera. Moreover, the analysis of the differences in optical density between duplicates indicated a high repeatability for the ELISA. At the ROC optimized cutoff, the sensitivity and specificity of the test were, respectively, 99.2% and 90.6% (specificity of 95.6% when excluding the samples from multiparasitized persons from Tanzania). The validated ELISA showed good performance in terms of sensitivity, repeatability, and reproducibility, whereas the specificity was limited. These results suggest that this test is suitable for detecting anti-Trichinella antibodies in human sera for diagnostic purposes, whereas its use in epidemiological surveys could be questionable.

PubMed Disclaimer

Figures

FIG. 1.

Scatter plot of the differences between the OD (recorded at 450 nm) of the serum duplicates and their means. Mean+2SD, mean plus two standard deviations; Mean−2SD, mean minus two standard deviations.

FIG. 2.

The ROC curve built for 1,159 serum samples from healthy blood donors and 367 samples from persons with confirmed trichinellosis. The AUC, which indicates accuracy, was determined for IE and for the mean of the OD duplicates and was equal to 0.999.

FIG. 3.

Western blot profiles obtained with sera from _Trichinella_-infected persons with different IgG levels by ELISA. A, molecular mass markers in kDa; lane 1, serum sample that yielded an OD of 2.0; lane 2, sample that yielded an OD of 1.5; lane 3, sample that yielded an OD of 0.5; lane 4, sample that yielded an OD of 0.4.

References

1. Akisu, C., S. B. Delibaçs, and S. Ozkoç. 2005. Evaluation of three different ELISA kits for the diagnosis of trichinellosis. Mikrobiyol. Bul. 39**:**325-331. - PubMed
1. Andiva, S., H. Yera, S. Haeghebaert, C. Tourte-Schaefer, J.-F. Magnaval, and J. Dupouy-Camet. 2002. Evaluation comparative d'un test d'agglutination au latex, de deux tests ELISA et d'un test western blot pour le diagnostic sérologique de la trichinellose humaine. Ann. Biol. Clin. (Paris) 60**:**79-83. - PubMed
1. Aronstein, W. S., S. A. Lewis, A. P. Norden, J. P. Dalton, and M. Strand. 1986. Molecular identity of a major antigen of Schistosoma mansoni which cross reacts with Trichinella spiralis and Fasciola hepatica. Parasitology 92**:**133-151. - PubMed
1. Au, A. C., R. C. Ko, J. W. Simon, N. J. Ridell, F. W. Wong, and M. J. Templer. 1983. Study of acute trichinosis in Ghurkas: specificity and sensitivity of enzyme-linked immunosorbent assays for IgM and IgE antibodies to Trichinella larval antigens in diagnosis. Trans. R. Soc. Trop. Med. Hyg. 77**:**412-415. - PubMed
1. Barennes, H., S. Sayasone, P. Odermatt, A. De Bruyne, S. Hongsakhone, P. N. Newton, O. Vongphrachanh, B. Martinez-Aussel, M. Strobel, and J. Dupouy-Camet. 2008. A major trichinellosis outbreak suggesting a high endemicity of Trichinella infection in northern Laos. Am. J. Trop. Med. Hyg. 78**:**40-44. - PMC - PubMed

Validation of an enzyme-linked immunosorbent assay for diagnosis of human trichinellosis - PubMed (original) (raw)