What blood tests should be done to work up rouleaux on a blood smear?

Medical Advisory BoardAll articles are reviewed for accuracy by our Medical Advisory Board
Educational purpose only • Exercise caution as content is pending human review
Article Review Status
Submitted
Under Review
Approved

Last updated: October 15, 2025View editorial policy

Personalize

Help us tailor your experience

Which best describes you? Your choice helps us use language that's most understandable for you.

Diagnostic Workup for Rouleaux Formation on Blood Smear

When rouleaux formation is identified on a peripheral blood smear, a focused workup for plasma cell disorders should be initiated, as multiple myeloma is the most clinically significant cause requiring prompt intervention. 1

Initial Laboratory Evaluation

  • Complete blood count with differential and peripheral blood smear examination to assess for additional findings such as circulating plasma cells, which may accompany rouleaux formation 2, 1
  • Comprehensive biochemistry panel including liver function tests, renal function tests, electrolytes, calcium, and albumin 2, 1

Specific Testing for Plasma Cell Disorders

  • Serum protein electrophoresis (SPEP) to detect and quantify monoclonal proteins 2, 1
  • Urine protein electrophoresis (UPEP) using a 24-hour urine collection (not random samples) 2, 1
  • Immunofixation electrophoresis of both serum and urine to confirm the presence and type of monoclonal protein, even if electrophoresis shows no measurable protein 2, 1
  • Serum free light chain assay to detect excess kappa or lambda light chains 1
  • Quantitative immunoglobulins by nephelometry, which complements electrophoresis and is particularly useful for measuring low levels of uninvolved immunoglobulins 2, 1

Further Evaluation if Monoclonal Protein is Detected

  • Bone marrow aspiration and biopsy to assess for clonal plasma cells 2, 1
  • Skeletal imaging (skeletal survey, low-dose whole-body CT, MRI, or PET-CT) to evaluate for bone lesions 1

Clinical Pearls and Pitfalls

  • Rouleaux formation appears microscopically as red blood cells lined up against each other, resembling "stacked coins" 3
  • Saline replacement can help distinguish true agglutination from rouleaux formation - true agglutination will remain when plasma is replaced with saline, while rouleaux will disappear when plasma proteins are removed 3
  • Nephelometric quantitation may overestimate monoclonal protein concentration when values are high, so both densitometer tracing and nephelometric quantitation are recommended 2
  • Never delay appropriate workup when rouleaux is identified, as it may be the first indication of a plasma cell dyscrasia requiring urgent intervention 1
  • While rouleaux can be caused by increased plasma proteins from conditions other than plasma cell disorders (such as inflammatory states), the diagnostic approach should prioritize ruling out multiple myeloma due to its significant morbidity and mortality implications 1

References

Guideline

Diagnostic Approach to Rouleaux Formation in Blood Smear

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Research

Rouleaux and saline replacement.

Immunohematology, 2018

Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

Have a follow-up question?

Our Medical A.I. is used by practicing medical doctors at top research institutions around the world. Ask any follow up question and get world-class guideline-backed answers instantly.