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. 2013 Jul;7(4):437-42.
doi: 10.5009/gnl.2013.7.4.437. Epub 2013 Jun 11.

Development of an ELISA for Quantitative Detection of Immunoglobulin G (IgG) and IgA Antibodies to Helicobacter pylori for Use in Korean Patients with H. pylori-Associated Diseases

Ji-Hyun Seo  1 , Jin-Su Jun, Hee-Shang Youn, Jung Sook Yeom, Ji Sook Park, Chan-Hoo Park, Hyang-Ok Woo, Woo-Kon Lee, Myung-Je Cho, Kwang-Ho Rhee
Affiliations

Development of an ELISA for Quantitative Detection of Immunoglobulin G (IgG) and IgA Antibodies to Helicobacter pylori for Use in Korean Patients with H. pylori-Associated Diseases

Ji-Hyun Seo et al. Gut Liver. 2013 Jul.

Abstract

Background/aims: We aimed to develop a quantitative enzyme-linked immunosorbent assay (ELISA) using whole-cell lysates of Helicobacter pylori 51 and to investigate its validity.

Methods: Data from 300 plates were obtained by two different operators. Standard sera were used to make a standard curve to analyze the quantity of anti-H. pylori immunoglobulin G (IgG) and IgA antibody. We obtained reproducible data with fewer dilutions of samples by the addition of serially diluted standard serum to each ELISA plate. To evaluate the validity of this ELISA, the 114 H. pylori-positive and -negative subjects were stratified into four age groups, i.e., 0 to 4, 5 to 9, 10 to 15, and 20 to 29 years, before testing.

Results: The mean IgG-antibody titers in H. pylori-positive and -negative subjects were 1,766.4 IU/mL and 654.3 IU/mL (p<0.001). The mean IgA-antibody titers in H. pylori-positive and -negative subjects were 350.1 IU/mL and 193.5 IU/mL (p<0.001). Anti-H. pylori IgG and IgA titers in the four age groups were higher in H. pylori-positive subjects than in H. pylori-negative subjects (p<0.05).

Conclusions: Using the current ELISA based on whole-cell lysates of H. pylori 51, reliable anti-H. pylori antibody titers were obtained regardless of the subject's age.

Keywords: Antibodies; Enzyme-linked immunosorbent assay; Helicobacter pylori.

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Conflict of interest statement

No potential conflict of interest relevant to this article was reported.

Figures

Fig. 1
Fig. 1
Comparison of enzyme-linked immunosorbent assay antibody units derived from single versus multiple serial dilutions. (A) Immunoglobulin G (IgG). (B) IgA. Antibody units were calculated from the OD492 reading of a single dilution referred to a standard curve and plotted on the X-axis. The standard sera were serially diluted to generate a complete dilution curve. The reciprocal of the dilution giving an OD492 reading of 1 was calculated (on the Y-axis) from the dilution curve. OD, optical density.
Fig. 2
Fig. 2
Quality-control parameters for an enzyme-linked immunosorbent assay standard curve. The R2 of a standard curve (fitted to a 4-parameter hyperbolic curve) is plotted on the X-axis, and the arithmetic mean value of OD492 of 4 blank wells on the same plate is plotted on the Y-axis. (A) Immunoglobulin G (IgG). (B) IgA. OD, optical density.
Fig. 3
Fig. 3
Comparison of immunoglobulin G (IgG) antibody titers in Helicobacter pylori (HP)-positive and H. pylori-negative groups according to age. The median antibody titer is higher in the H. pylori-positive group than in the H. pylori-negative group. *p<0.0001.
Fig. 4
Fig. 4
Comparison of immunoglobulin A (IgA) antibody titers in the Helicobacter pylori (HP)-positive and H. pylori-negative groups according to age. The median antibody titer is higher in the H. pylori-positive group than in the H. pylori-negative group. *p<0.05.
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