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Related Experiment Video

Updated: Jan 20, 2026

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Calreticulin Mutation by Immunohistochemistry: Can It Replace PCR?

Aastha Gupta1, Sudha Sazawal, Manoranjan Mahapatra

  • 1Department of Hematology, All India Institute of Medical Sciences, New Delhi, India.

Applied Immunohistochemistry & Molecular Morphology : AIMM
|September 4, 2019
PubMed
Summary

Immunohistochemistry (IHC) effectively detects Calreticulin (CALR) mutations in myeloproliferative neoplasms. This simple, cost-effective method shows high accuracy, making it ideal for resource-limited settings.

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Area of Science:

  • Hematology
  • Molecular Pathology
  • Oncology

Background:

  • Primary myelofibrosis (PMF) and essential thrombocythemia (ET) diagnosis was challenging before 2013 due to a lack of reliable clonal markers.
  • Calreticulin (CALR) mutations were identified in 60-88% of JAK2/MPL-negative PMF and ET cases, serving as a crucial diagnostic marker.

Purpose of the Study:

  • To evaluate the utility of immunohistochemistry (IHC) for detecting CALR mutations.
  • To compare the diagnostic performance of IHC with polymerase chain reaction (PCR) for CALR mutation detection in JAK2-negative PMF and ET.

Main Methods:

  • Thirty-one JAK2V617F-negative PMF and ET cases were analyzed.
  • CALR mutation status was determined using both PCR and IHC with an antimutated CALR antibody (CAL2).
  • IHC performance was assessed by calculating sensitivity, specificity, positive predictive value, and negative predictive value against PCR results.

Main Results:

  • CALR mutations were detected in 64.5% (20/31) of the evaluated cases.
  • The prevalence of CALR mutations was 60.9% in JAK2-negative PMF and 75% in ET.
  • IHC demonstrated high accuracy with 89.4% sensitivity, 100% specificity, and excellent agreement (κ=0.86) with PCR.

Conclusions:

  • Immunohistochemistry (IHC) is a highly sensitive and specific method for detecting CALR mutations in PMF and ET.
  • IHC is a cost-effective and accessible alternative to molecular methods, particularly beneficial for resource-limited settings.
  • IHC serves as an excellent screening test for CALR mutations in JAK2-negative myeloproliferative neoplasms.