Utility of CD157 as a Common Leukocytes Marker for Paroxysmal Nocturnal Hemoglobinuria Screening in a Single Tube Five Color Combination
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To study the utility and advantage of CD157 in the paroxysmal nocturnal hemoglobinuria (PNH) screening along with its ability to replace CD24 and CD14. This was a confirmatory study to analyse the role and advantage of CD157 in a single tube five color combination to identify the PNH clones. A serial tenfold dilution experiments was carried out for sensitivity assessment. Reproducibility was checked in the intra-assay and inter-assay experiments. The results obtained with CD157 based assay were compared with the routinely used single tube six color CD24/CD14 based assay. CD157 showed a high degree of sensitivity at the level of 10−4. PNH positive clone sizes were precise with CVs of inter-assay and intra-assay precision analysis for polymorphs/monocytes ranging from 2.94 to 4.31/2.52 to 8.93, and 0.91 to 3.23/1.65 to 5.33%; respectively. The results were similar to those obtained from CD24/CD14 based assay (R2 > 0.993). There was no false positive or false negative result. CD157 was found better in delineating the type II clones. CD157 can be used as a common PNH leucocyte marker with high degree of sensitivity and precision. It can replace CD24 and CD14 from the currently used assays and thus bring down the cost of PNH screening.
KeywordsCD157 Flow cytometry (FCM) Paroxysmal nocturnal hemoglobinuria (PNH)
The authors would like to thanks Mr Kaushal Kumar, Technical staff, Flow cytometry Lab, SGPGI, Lucknow.
Compliance with Ethical Standards
Conflict of interest
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Informed consent was taken from all the individuals participants included in this study.
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