TY - JOUR
T1 - 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
AU - Aziz, Faisal
AU - Sherwani, Sikander Khan
AU - Akhtar, Syed Shakeel
AU - Kazmi, Shahana Urooj
PY - 2014/1
Y1 - 2014/1
N2 - 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.
AB - 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.
KW - Enzyme-linked immunoassay
KW - Gastroduodenal ulcer disease
KW - H. pylori surface whole cell antigen
KW - Helicobacter pylori
UR - http://www.scopus.com/inward/record.url?scp=84891661449&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84891661449&partnerID=8YFLogxK
U2 - 10.1007/s11274-013-1448-4
DO - 10.1007/s11274-013-1448-4
M3 - Article
C2 - 23921679
AN - SCOPUS:84891661449
SN - 0959-3993
VL - 30
SP - 305
EP - 315
JO - World Journal of Microbiology and Biotechnology
JF - World Journal of Microbiology and Biotechnology
IS - 1
ER -