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Suman Thomer

Surgeon, MO Civil Hospital Rohtak, India

Title: FLOW CYTOMETRIC DETECTION OF CD64 ON NEUTROPHILS AND HLA-DR ON MONOCYTES AS MARKERS OF NEONATAL SEPSIS

Biography

Biography: Suman Thomer

Abstract

INTRODUCTION

Neonatal sepsis remains a diagnostic burden globally responsible for about 30-50% of the total neonatal deaths each year in developing countries. Neutrophil CD64 is found to be a promising marker for diagnosis of early and late infections in newborns. Decreased expression of HLA-DR on monocytes has been associated with decreased survival in newborn with sepsis. The purpose of this study is to evaluate the role of neutrophil CD64 and monocyte HLA-DR expression in diagnosis and prognosis of neonatal sepsis.

MATERIALS AND METHODS

This was a prospective study carried out in Department of Pathology in collaboration with Neonatal services division, Department of Paediatrics, Pt.B.D.Sharma Post Graduate Institute of Medical Sciences, Rohtak. Seventy full term  neonates with clinical suspicion of sepsis and warranting antibiotic therapy were enrolled provided they had not received antibiotics in the preceding 72 hours after an informed parental consent. Thirty age matched healthy neonates were taken as controls. The peripheral venous blood sample was collected for flow cytometry, blood culture and sepsis screening in all patients. Cell surface markers (CD64 on neutrophils and HLA-DR on monocytes) expression was measured by 8 Color Flow cytometer BD FACS Canto II (Becton Dickinson, San Jose,CA) system. A composite parameter was derived by division of MFI (mean fluorescence intensity) values of nCD64 and their respective mHLA-DR and multiplying the ratio by 100, and termed as sepsis index (Sepsis Index= nCD64/mHLA-DR x 100).

 RESULTS

Out of the 70 neonates studied nineteen cases were diagnosed as definite sepsis (culture positive), 27 as probable sepsis (sepsis screen positive but culture negative) and 24 as no sepsis. nCD64 positivity was seen in all cases of definite sepsis and 88.89% cases of probable sepsis. mHLA DR was positive in 63.16% cases of definite sepsis, 85.19% in probable sepsis and 54.17% of no sepsis group. Out of 19 cases of definite sepsis 94.74% cases had positive sepsis index while one case had sepsis index less than cutoff value. In our study, nCD64 revealed 100% sensitivity, 87.5% specificity, 86.36% positive predictive value, 100 % negative predictive value and 93.02% of diagnostic accuracy in culture positive sepsis. However; downregulation of mHLA-DR observed in our study alone showed a poor diagnostic utility. In definite sepsis group mHLA DR revealed sensitivity of 63.15%, specificity of 45.83%, PPV of 48%, NPV of 61.11% and accuracy of 53.48%. Sepsis index was calculated in all the neonates in this study and found sensitivity of 94.73%, specificity of 62.50%, PPV of 66.66% and NPV of 93.75% and accuracy of 76.74% in definite sepsis group