Development of an in-house enzyme-linked immunosorbent assay based on surface whole cell antigen for diagnosis of Helicobacter pylori infection in patients with gastroduodenal ulcer disease

Faisal Aziz, Sikander Khan Sherwani, Syed Shakeel Akhtar, Shahana Urooj Kazmi

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Helicobacter pylori (H. pylori) is a causative agent of gastritis, gastroduodenal ulcers and gastric adenocarcinoma. More than 50 % world population is colonized by H. pylori, which is closely related to the chronic gastritis and gastric ulcer infection. In this study, a total of 214 gastritis patient's serum samples were screened for anti-H. pylori IgG antibody. A 96-well plate coated with 20 μg/ml antigen and hundred-fold diluted patient's serum was allowed to react. After extensive washing with buffer, 1:2,500 diluted conjugated secondary antibody was added. Later substrate was added to observe positivity by measuring the intensity of color. Statistical analyses were performed, and p value of <0.01 was taken as significant; 84 % male patients and 89 % female patients, respectively, tested positive for H. pylori, while agewise distribution was 35-45 years males (40 %) and 35-55 years females (52 %) were found highest number of H. pylori infected patients. In-house ELISA based on surface whole cell antigen (wELISA) showed a sensitivity of 93 %, specificity of 100 %, accuracy 94 % and κ value 0.86 with significant correlation R-0.77020; p < 0.0001. We conclude that H. pylori local isolates surface antigen was satisfactory for diagnosis as different parameters were adjusted according to the local H. pylori isolates. Fluctuations in serum antibody titer predict the variation in an individual's response of the host against H. pylori. In-house wELISA could provide a reliable and a clinically useful method for the diagnosis of H. pylori infection in patients of Karachi, Pakistan.

Original languageEnglish (US)
Pages (from-to)305-315
Number of pages11
JournalWorld Journal of Microbiology and Biotechnology
Volume30
Issue number1
DOIs
StatePublished - Jan 1 2014
Externally publishedYes

Keywords

  • Enzyme-linked immunoassay
  • Gastroduodenal ulcer disease
  • H. pylori surface whole cell antigen
  • Helicobacter pylori

Fingerprint

Dive into the research topics of 'Development of an in-house enzyme-linked immunosorbent assay based on surface whole cell antigen for diagnosis of Helicobacter pylori infection in patients with gastroduodenal ulcer disease'. Together they form a unique fingerprint.

Cite this