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<p>Fetomaternal hemorrhage (FMH) involves the transfer of fetal red blood cells into the maternal circulation, posing a risk of maternal alloimmunization against fetal red cell antigens, particularly RhD. Inadequate quantification may lead to incorrect dosing of Rh immunoglobulin (RhIG), increasing the risk of hemolytic disease of the fetus and newborn (HDFN). Accurate and accessible diagnostic methods are therefore essential, especially in resource-limited settings. This study aimed to quantify FMH using two diagnostic modalities—the Kleihauer–Betke test (KBT) and flow cytometry (FCM)—and to compare their diagnostic performance in a tertiary care setting in India. A cross-sectional comparative study was conducted from May 2024 to May 2025. Maternal blood samples from 50 term-delivering women were analyzed using both KBT, following the standard acid-elution technique, and flow cytometry with dual staining for fetal hemoglobin and carbonic anhydrase. FMH volume was calculated assuming a maternal blood volume of 5,000 mL. Significant FMH (&gt; 30 mL) was detected in 28 cases (56%) by KBT and in 23 cases (46%) by flow cytometry. A very strong positive correlation was observed between the two methods (<i>r</i> = 0.986; <i>p</i> &lt; 0.001). KBT demonstrated 100% sensitivity and 81.5% specificity when flow cytometry was used as the reference standard. Although FCM remains the most accurate method for FMH quantification, KBT shows excellent correlation and high diagnostic reliability. In resource-constrained environments, KBT serves as a practical, cost-effective, and accessible tool for guiding RhIg prophylaxis, thereby aiding in the prevention of alloimmunization-related complications.</p>

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Real-World Utility Of The Kleihauer–Betke Test For Fetomaternal hemorrhage-Guided RhIG Prophylaxis In Indian Tertiary Care: A Comparative Study Against Flow Cytometry In Term Deliveries

  • 期刊论文

  • 发表日期：

    2026年6月15日

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  • Anurag Gairola,
  • Ujjwal Dimri,
  • Anoop Sharma,
  • Akshaya Tomar,
  • Chinmay Shrikrishna Pendharkar,
  • Sujay Bhowmik

摘要

Fetomaternal hemorrhage (FMH) involves the transfer of fetal red blood cells into the maternal circulation, posing a risk of maternal alloimmunization against fetal red cell antigens, particularly RhD. Inadequate quantification may lead to incorrect dosing of Rh immunoglobulin (RhIG), increasing the risk of hemolytic disease of the fetus and newborn (HDFN). Accurate and accessible diagnostic methods are therefore essential, especially in resource-limited settings. This study aimed to quantify FMH using two diagnostic modalities—the Kleihauer–Betke test (KBT) and flow cytometry (FCM)—and to compare their diagnostic performance in a tertiary care setting in India. A cross-sectional comparative study was conducted from May 2024 to May 2025. Maternal blood samples from 50 term-delivering women were analyzed using both KBT, following the standard acid-elution technique, and flow cytometry with dual staining for fetal hemoglobin and carbonic anhydrase. FMH volume was calculated assuming a maternal blood volume of 5,000 mL. Significant FMH (> 30 mL) was detected in 28 cases (56%) by KBT and in 23 cases (46%) by flow cytometry. A very strong positive correlation was observed between the two methods (r = 0.986; p < 0.001). KBT demonstrated 100% sensitivity and 81.5% specificity when flow cytometry was used as the reference standard. Although FCM remains the most accurate method for FMH quantification, KBT shows excellent correlation and high diagnostic reliability. In resource-constrained environments, KBT serves as a practical, cost-effective, and accessible tool for guiding RhIg prophylaxis, thereby aiding in the prevention of alloimmunization-related complications.