Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Detection of cell specific cluster determinant expression by reverse transcriptase polymerase chain reaction

M James-Yarish1, W G Bradley, P J Emmanuel

  • 1Department of Pediatrics, All Children's Hospital, University of South Florida, St. Petersburg 33701.

Journal of Immunological Methods
|February 28, 1994
PubMed
Summary

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Identification of a novel allele HLA-DQB1*0622.

Tissue antigens·2007
Same author

Characterization of a novel allele HLA-B*3549.

Tissue antigens·2005
Same author

Bone metabolism in children with human immunodeficiency virus infection receiving highly active anti-retroviral therapy including a protease inhibitor.

The Journal of pediatrics·2001
Same author

Radiological case of the month. Aneurysmal dilation of cerebral arteries associated with HIV infection.

Archives of pediatrics & adolescent medicine·2001
Same author

A retroviral-derived immunosuppressive peptide activates mitogen-activated protein kinases.

Journal of immunology (Baltimore, Md. : 1950)·2001
Same author

Immunologic reconstitution following bone marrow transplantation for X-linked hyper IgM syndrome.

Clinical immunology (Orlando, Fla.)·2001

A new reverse transcriptase polymerase chain reaction (RT-PCR) method offers sensitive gene expression analysis for cell surface markers. This technique detected B cell markers in a patient previously thought to lack them, improving diagnostic capabilities.

Area of Science:

  • Molecular Biology
  • Immunology
  • Biochemistry

Background:

  • Flow cytometry is commonly used for detecting cell surface markers.
  • Gene expression analysis of cell determinants is crucial in immunology.
  • Existing methods may lack sensitivity for certain cell populations.

Purpose of the Study:

  • To develop a novel, sensitive, and reproducible method for analyzing gene expression of cell surface determinants.
  • To compare the new method with traditional flow cytometry.
  • To demonstrate the utility of the new method in a clinical case.

Main Methods:

  • Utilized reverse transcriptase polymerase chain reaction (RT-PCR) for gene expression analysis.
  • Quantified PCR products using radiolabeled nucleotides and ion exchange filter chromatography.

Related Experiment Videos

  • Applied the method to human cell lines, peripheral blood lymphocytes, bone marrow, and lymph node cells.

    • Main Results:

    • The RT-PCR method demonstrated high sensitivity and reproducibility for analyzing CD gene expression.
    • Analysis of a B cell deficient patient revealed CD19, CD20, and CD23 expression via RT-PCR, which was not detected by flow cytometry.
    • Results from RT-PCR and flow cytometry were largely similar, but RT-PCR offered enhanced detection capabilities.

    Conclusions:

    • RT-PCR provides a sensitive alternative for detecting cell surface marker gene expression.
    • This method can identify cell populations missed by conventional techniques, aiding in diagnosing immune deficiencies.
    • The described methodology enhances the understanding of gene expression in various human cell types.