Rh-Negative Female with Anti-D at Delivery: A Case Study

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Author: Pat Letendre, MEd
Reviewer: Erin Tretter, MT(ASCP)

If you have completed the course, Hemolytic Disease of the Fetus and Newborn, you will enjoy working through a case that provides real-world application of what you have learned.

This case study presents an opportunity to review perinatal testing programs and the crucial role of RhIg in preventing hemolytic disease of the fetus and newborn (HDFN) due to anti-D. The case also examines practical aspects of routine serologic testing involving neonates and women who have received RhIg during pregnancy.

You will be guided through laboratory findings that need to be interpreted and resolved; be presented with current best practices in perinatal testing programs; review the characteristics of RhIg and its use in pregnancy; review and investigate key issues associated with detection of anti-D in women who have received antenatal RhIg; and learn about crossmatch and LIS policies related to RhIg-derived passive anti-D.

Continuing Education Credits


  • Discuss the history of RhIG, its sources, preparation, constituents, safety, types, mechanisms of action, criteria for the administration of RhIG antenatally and postnatally, dosage calculation, and causes of failures.
  • Interpret serologic tests done on the mother, father, and fetus / newborn as part of perinatal testing programs, including antibody identification.
  • Identify best practices for serologic and other tests used in perinatal testing programs.
  • Describe the principles, uses, and limitations of the rosette test, Kleihauer-Betke test, and flow cytometry in Rh prevention programs.
  • Discuss and resolve key issues associated with detection of anti-D in women who have received antenatal RhIG.
  • Identify crossmatch and LIS policies and practices related to RhIG-derived passive anti-D.

Course Outline

  • Case Scenario
      • Introduction
      • Case Presentation
    • Initial Serologic Test Results
      • ABO, Rh, and Antibody Screen
      • Which of the following are possible causes of the positive antibody screen?
      • Why might screen cell #2 be reacting stronger than screen cell #1?
      • Antigram to Explain Prior Question
      • Which of the possible causes is the MOST likely cause of the positive antibody screen?
  • Perinatal Testing Programs
      • Perinatal Testing Programs
      • Immunogenicity
      • Routine Serologic Tests: Mother
      • Routine Serologic Tests: Newborn Protocols
      • Routine Serologic Tests: Newborn Protocols, continued
  • Interpreting Initial Serologic Tests
      • Interpreting Initial Serologic Tests
      • Antibody Exclusion Protocol (General)
      • Initial Antibody Exclusions
      • Antibody Exclusion Protocols (RhIG)
      • Passive or Active Immunization?
      • RhIG-Derived Anti-D Reaction Strength
      • Factors Affecting RhIg Reaction Strength
      • How Long Can RhIG Be Detected?
      • Which of the following factors is most likely to cause a false-negative antibody screen following RhIG injection?
      • A pregnant female has been injected with RhIG antenatally and has a positive antibody screen at delivery. If the antibody has been confirmed as anti-D...
      • A pregnant female who received RhIG at 28 weeks gestation has a positive antibody screen at delivery. If the antibody has been confirmed as anti-D alo...
  • Follow-up Serologic Tests
      • How Much Testing?
      • Mini-Panel Antibody Exclusion
      • Using the guidelines in the antibody exclusion protocol, all unexcluded antibodies (anti-C, E, K, Fyb, Jka, M, s, Leb) have been excluded by the mini-...
  • Post-delivery Testing
      • Serologic Tests on Newborn
      • The newborn's ABO group is invalid because no reverse (serum) group was set up with A1 and B cells.
      • The newborn's Rh(D) type is invalid because the DAT is positive.
      • The positive DAT on the newborn means that the infant probably has clinically significant hemolysis.
      • Newborn's Clinical Status
      • Is the mother a candidate for RhIG? (type Y for yes or N for no)
      • Screening for Fetomaternal Hemorrhage (FMH)
      • Quantifying FMH
      • Kleihauer-Betke (KB) Test
      • Flow Cytometry
      • Calculating RhIG Dosage
      • Assessing FMH and RhIG Dosage
      • Which of the following tests are suitable for quantifying the size of fetomaternal hemorrhage (FMH)? Select all that apply.
      • Facilities that use a special calculation for RhIG dosage add one vial, regardless if they round up or round down.
  • Summary and Conclusions
      • Case Summary
  • When Transfusion Is Necessary
      • Crossmatch Issues
      • Crossmatch Implications of RhIg-associated Passive Anti-D
      • Crossmatch Practices Related to RhIG - Introduction
      • LIS Issues Related to RhIG
      • Crossmatch Practices Related to RhIG: Examples
      • Blood safety standards such as AABB Standards directly specify that an electronic crossmatch cannot be done when an Rh negative female has an anti-D c...
  • Further Reading
      • Further Reading
  • References
      • References

Additional Information

Level of instruction: Intermediate

Intended Audience: Clinical laboratory technologists, technicians, and pathologists. This course is also appropriate for clinical laboratory science students and pathology residents.
Author information: Pat Letendre, MEd is a laboratory technologist, educator, and consultant. Currently, she consults full-time in the areas of transfusion medicine, education, professional development, and use of the Internet in education. Ms. Letendre is the Webmaster for Canada's transfusion safety officers and the TraQ website coordinator. She holds a Masters of Education degree in adult education from the University of Alberta and a Bachelor of Science degree from the University of Manitoba.  
Reviewer information: Erin Tretter, MT(ASCP), is currently the STAT Laboratory Supervisor at Penn Presbyterian Medical Center in Philadelphia, PA. She received her BS in Medical Technology from California University of Pennsylvania and has nearly 8 years of experience as a Generalist, including Blood Bank, Hematology and Chemistry. Erin is the Blood Bank Clinical Instructor for the Clinical Laboratory Science Program at St. Christopher’s and has 4 years experience teaching immunohematology concepts and laboratory procedures to Medical Technology students. She has also provided blood bank training for laboratory technologists and medical students. Erin is currently obtaining a Master’s in Business Administration from Florida Institute of Technology where she is a member of the Phi Kappa Phi Honor’s Society.

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This course is part of:
Antibody screen cells