Preparation of colloidal gold immunochromatographic strip for detection of Paragonimiasis skrjabini

PLoS One. 2014 Mar 18;9(3):e92034. doi: 10.1371/journal.pone.0092034. eCollection 2014.

Abstract

Background: Paragonimiasis is a food-borne trematodiasis, a serious public health issue and a neglected tropical disease. Paragonimus skrjabini is a unique species found in China. Unlike paragonimiasis westermani, it is nearly impossible to make a definitive diagnosis for paragonimiasis skrjabini by finding eggs in sputum or feces. Immunodiagnosis is the best choice to detect paragonimiasis skrjabini. There is an urgent need to develop a novel, rapid and simple immunoassay for large-scale screening patients in endemic areas.

Methodology/principal findings: To develop a rapid, simple immunodiagnostic assay for paragonimiasis, rabbit anti-human IgG was conjugated to colloidal gold particles and used to detect antibodies in the sera of paragonimiasis patients. The synthesis and identification of colloidal gold particles and antibody-colloidal gold conjugates were performed. The size of colloidal gold particles was examined using a transmission electron microscope (TEM). The average diameter of colloidal gold particles was 17.46 nm with a range of 14.32-21.80 nm according to the TEM images. The formation of antibody-colloidal gold conjugates was monitored by UV/Vis spectroscopy. Excretory-secretory (ES) antigen of Paragonimus skrjabini was coated on nitrocellulose membrane as the capture line. Recombinant Staphylococcus protein A was used to prepare the control line. This rapid gold immunochromatographic strip was assembled in regular sequence through different accessories sticked on PVC board. The relative sensitivity and specificity of the strip was 94.4% (51/54) and 94.1% (32/34) respectively using ELISA as the standard method. Its stability and reproducibility were quite excellent after storage of the strip at 4°C for 6 months.

Conclusions/significance: Immunochromatographic strip prepared in this study can be used in a rapid one-step immunochromatographic assay, which is instantaneous and convenient.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Helminth / chemistry*
  • Antibodies, Helminth / immunology
  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / immunology
  • Chromatography, Affinity / methods*
  • Chromatography, Affinity / standards
  • Colloids
  • Female
  • Gold / chemistry
  • Humans
  • Paragonimiasis / diagnosis*
  • Paragonimiasis / immunology
  • Paragonimiasis / parasitology
  • Paragonimus / immunology
  • Paragonimus / isolation & purification*
  • Particle Size
  • Rabbits
  • Reagent Strips
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Antibodies, Helminth
  • Antibodies, Monoclonal
  • Colloids
  • Reagent Strips
  • Gold

Grants and funding

This study was supported by the Natural Science Foundation of Chongqing (CSTC, 2007BB5055), the Military Pre-research Fund (No. 2009XYY01) and the Education Sciences Foundation of Sichuan (No. 2005A067). The author's laboratory was supported in part by grants from the National Natural Science Foundation of China (No. 30872366 and No. 30371261). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.