Pediatr Gastroenterol Hepatol Nutr.  2012 Jun;15(2):85-90.

Comparison of Four Commercial ELISA Kits and In-House Immunoblotting for Diagnosis of Helicobacter pylori Infection

• Jeong HL
• Jung YS
• Jun JS
• Yeom JS
• Park JS
• Seo JH
• Lim JY
• Park CH
• Woo HO
• Youn HS
• Ko GH
• Baik SC
• Lee WK
• Cho MJ
• Rhee KH

Abstract

PURPOSE
Commercial enzyme-linked immunosorbent assay (ELISA) kits have been considered less reliable for children than for adults. The aim of this study was to compare four ELISA kits and in-house immunoblotting based on the analysis of anti-H. pylori-IgG antibody reactivity.
METHODS
A total of 399 serum samples were collected at the GNU Hospital during 1998-1999. All sera were tested using ELISA and immunoblotting. Statistically significant differences were determined by the chi2 test.
RESULTS
The overall seropositivity rates using GAP IgG, Genedia IgG, HM-CAP, Pyloriset EIA-G, and immunoblotting were 13.0%, 25.1%, 18.3%, 15.8%, and 62.9%, respectively. Immunoblotting showed a higher seropositivity rate than did all four ELISA kits in all age groups. Genedia IgG had the highest seropositivity among the ELISA kits. The seropositivity rate for children aged 13 to 18 months was lowest, and that of children aged 15 years was highest (90.0%). The seropositivity rate for children aged 7 months to 5 years was significantly lower than that for children aged 6 to 15 years among the four ELISA kits (p<0.0001) and immunoblotting (p=0.02).
CONCLUSION
Immunoblotting is the most sensitive test for detection of anti-Helicobacter pylori IgG antibodies among the serological tests in this study. These results emphasize the need for standardization when commercial ELISA tests are used in different nations or in young age groups. Immunoblotting could be a suitable noninvasive assay for serodiagnosis and seroepidemiologic study of H. pylori infection in Korean children.

Keyword

ELISA; Immunoblot; Helicobacter pylori infection; Children

MeSH Terms

Adult
Aged
Antibodies
Child
Enzyme-Linked Immunosorbent Assay
Helicobacter
Helicobacter pylori
Humans
Immunoblotting
Immunoglobulin G
Seroepidemiologic Studies
Serologic Tests
Antibodies
Immunoglobulin G

Figure

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  Fig. 1 Immunoblot assay results were classified into three patterns based on immunoreactive bands. Only Pattern I, which shows reactivities against 120-kDa antigens as well as other antigens of H. pylori, was considered to be a specific marker of H. pylori infection in this study. Panel A shows a Ponceau S-stained nitrocellulose membrane onto which marker proteins and separated H. pylori antigen were transferred.

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  Fig. 2 Seropositivity rates of the four commercial ELISA kits and immunoblotting according to age. The seropositivity rates increased with age. The seropositivity rates of immunoblotting were higher than those of the ELISA kits, and the discrepancy in the seropositivity rates of anti-H. pylori IgG antibody was highest in 0- to 6-month-old infants.

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  Fig. 3 Comparisons of seropositivity rates for GAP IgG, Genedia IgG, HM-CAP, Pyloriset EIA-G, and immunoblotting in children 7 months to 5 years of age and 6 to 15 years of age. The seropositivity rates in children aged 6 years and older were higher than those of children aged 7 months to 5 years of age in all ELISA kits and immunoblotting. *p<0.0001, **p=0.02.

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  Fig. 4 Proportion of immunoblot patterns in the seropositive cases of the four ELISA kits. More than 80% were Pattern I. There were no results showing Pattern III.

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