This five-day workshop included wet-lab activities as well as lectures and group discussions. The workshop highlighted molecular assays for a variety of newborn disorders including galactosemia, hemoglobinopathies, CF and SCID. Some of the topics discussed included molecular laboratory design and unidirectional workflow, comparison of instrument platforms for NBS genotyping, assay requirements for multiplex assay design and data reporting and clinical interpretation. Laboratory activities included DNA extraction using the boil prep method, using robotics for DNA extraction and assay set-up, running and analyzing gels and DNA quantitation using RNAseP.
The presentations below have been authorized to be posted on this website. Duplication is not authorized without the express written permission of the authors.
Sponsors
Association of Public Health Laboratories
Centers for Disease Control and Prevention, Newborn Screening and Molecular Biology Branch
Co-Directors
Suzanne Cordovado, Centers for Disease Control and Prevention
Rachel Lee, Texas Department of State Health Services
Course Objectives
- Identify scenarios when a molecular test is appropriate in the newborn screening laboratory
- Describe the benefits of using a molecular test for different newborn screening disorders
- Name at least two applications of molecular testing in newborn screening
- Describe the common protocols for DNA extraction and DNA quantification from dried blood spots
- Describe a PCR-based assay to detect a single or small number of mutations
- Conduct DNA extraction, DNA quantification, PCR amplification, gel electrophoresis, and mutation detection methods
- Understand quality control procedures and requirements specific for molecular testing
- Understand data reporting and interpretation as it relates to molecular testing
Monday, March 9
Overview of NBS Molecular Assays
-Michele Caggana, NYS Department of Health
Overview of Hemoglobinopathies and Galactosemia
-Mike Glass, Washington State Department of Health
DNA Extraction and Quantification
-Suzanne Cordovado, Centers for Disease Control and Prevention
ARMS PCR Overview
-Rachel Lee, Texas State Department of State Health Services
Tuesday, March 10
Case Study of Molecular Assay Validation: Primary Molecular Screening
-Mei Baker, University of Wisconsin-Madison
Lab Math- Molarity
-Carlos Saavedra-Matiz, NYS Department of Health
Wednesday, March 11
Molecular Methods Used in NBS Genotyping
-Carlos Saavedra-Matiz, NYS Department of Health
Overview of Cystic Fibrosis
-Marie Earley and Suzanne Cordovado, Centers for Disease Control and Prevention
Case Study of Molecular Assay Validation 2nd Tier Molecular Screening
-Rachel Lee, Texas State Department of State Health Services
Lab Math- Primer Preparation and Dilutions
-Carlos, Saavedra-Matiz, NYS Department of Health
Post-Analytic Issues- Data Reporting and Clinical Interpretation
-Anne Comeau, University of Massachusetts Medical School
Thursday, March 12
Molecular Lab Design, QA/QC Considerations
-Rachel Lee, Texas State Department of State Health Services
Quality Assurance Program for Cystic Fibrosis Newborn Screening
-Marie Earley, Centers for Disease Control and Prevention
Newborn Screening for Severe Combined Immunodeficiency
-Jennifer Taylor, Centers for Disease Control and Prevention
Friday, March 13
Molecular Assessment Program
-Christopher Greene, Centers for Disease Control and Prevention
NBS Molecular Resources Website-Laura Hancock, Centers for Disease Control and Prevention
Course Exam