Microdetectives

Microdetectives
A Study of Humoral Response
Middle Level: Life Sciences
Lab Activity: "A Sandwich Named ELISA"

Abstract
A Secondary Science Classroom Laboratory Exercise Using the ELISA Assay for the Detection of Pathogen-Specific Humoral Response.

This lesson was designed to introduce students to basic immunological concepts through formal, scientific research, stimulate inquiry regarding primary and secondary immune response, apply the ELISA assay results to selected pathogens, and reinforce public health issues corresponding with their laboratory experience. Students were issued plates previously coated with anti-mouse Ig. Vials of serum were distributed for titration and then micropipetted into plate wells. Plates underwent refrigeration at 4^oC overnight. Plates were then washed and conjugated anti-human Ig (w/ horseradish perioxidase) was added to each well. The samples underwent an incubation period at 37^oC for 30-60 minutes. Plates were washed again and substrate was added. Results were monitored visually by color change. Parallel plates were developed at the NYS Health Department Cellular Immunology Laboratory to permit quantitation utilizing the ELISA Plate Reader. Data analysis was performed by students. Class discussions referred to ELISA results to assess exposure to pathogen. Laboratory equipment was acquired through the Greater Capital Region DNA Science Project, Albany, NY.

Overview
In this curriculum unit, students are presented with a mystery scenario involving two friends who are exposed to the same pathogen, but one recovers without medical attention. Students are guided through this scenario by performing an ELISA analysis and applying their understanding of humoral (immune) response to their experimental results.

contexts in which it could be used in a biology course

To reinforce units from the NYS Biology syllabus:
Structure and study of cells/cell biochemistry/human body systems/mitosis: cancer /evolution of viruses and bacteria/genetics

general purpose of the laboratory unit

To introduce students to basic immunological concepts through formal, scientific research, stimulate inquiry regarding primary and secondary immune response.

To apply the ELISA assay results to selected pathogens, and reinforce public health issues corresponding with their laboratory experience.

what students will be doing/technical skills that will be acquired

Students will use micropipettes to titrate and then aliquot serum into 96-well plates (previously coated with anti-mouse Ig). Immediate enzyme/substrate reactions will be visually observed. Quantitative results from the parallel plates will be processed at Wadsworth laboratories using an ELISA plate reader. This data can then be graphed and analyzed by the students.

relevancy to student learning/lives

This unit includes health issues which concern future generations. Viral and bacterial infections plague younger people and their environment each year. The human immunodeficiency virus, the Ebola virus, the yearly mutating influenza viruses, antibiotic-resistant strains of common bacteria (i.e. streptococci), and carcinogenic substances, all initiate discussion amongst the youth of today. This unit is designed to provide the students with the opportunity to discuss issues relevant to their generation.

To receive a copy of this curriculum, please contact: The American Association of Immunologists, telephone: (301) 634-7178, fax: (301) 634-7887, email infoaai@aai.org.