Luminex technology for anti‐HLA antibody screening: Evaluation of performance and of impact on laboratory routine

Colombo, M. B.; Haworth, S.; Poli, Francesca; Nocco, Angela; Puglisi, Giuseppe; Innocente, Annalisa; Serafini, Marta; Messa, Piergiorgio; Scalamogna, M.

doi:10.1002/cyto.b.20353

Cited by 95 publications

(72 citation statements)

References 16 publications

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“…(ii) Luminex method: Two lasers are used to excite the fluorochrome of the bead and the phycoerythrin bound to the antibody. The degree of fluorescence is calculated as a mean fluorescence intensity (MFI) [13].…”

Section: Quantitation Of the Resultsmentioning

confidence: 99%

An Update on Crossmatch Techniques in Transplantation

Kumar¹,

Mohiuddin²,

Sharma³

et al. 2017

J Kidney

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Crossmatch techniques formulated nearly a half century ago remain the bedrock of transplantation science. While initial methods relied on complement dependent cytotoxicity. More recently additions to the transplant physician's immunologic armamentarium have arrived in both cell-based as well as solid phase assays. Flow cytometry is useful in previously sensitized patients while bead technology has revolutionised HLA antibody testing. The virtual crossmatch uses bead technology to detect specific recipient antibodies without requiring donor serum. We present a concise review of the techniques available, their basis and comparison between them so as to better understand their respective applications.

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Section: Quantitation Of the Resultsmentioning

confidence: 99%

An Update on Crossmatch Techniques in Transplantation

Kumar¹,

Mohiuddin²,

Sharma³

et al. 2017

J Kidney

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“…17 Traditionally, the status of sensitization of patients has been defined by the CDC method; however, this method has shown some limitations due to the panel composition, the quality of the fresh cells used, and the difficulty to distinguish class II antibodies in the presence of antibodies of class I. 18,19 Conversely, new methodologies such as the Luminex platform have demonstrated both a greater sensitivity and an unambiguous recognition of antibodies against one or both classes. 20 The use of bead-based Luminex screening in laboratories has become widespread worldwide and has improved the detection of anti-HLA antibodies.…”

Section: Discussionmentioning

confidence: 99%

Untitled

Minucci

Resse

Sabia³

et al. 2017

Exp Clin Transplant

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Objectives: For decades, the detection of anti-HLA antibodies in candidates for solid-organ transplant has been performed with the traditional complementdependent cytotoxicity method; this assay has been then integrated with the introduction of solid-phase assays. Over the past 20 years, the Luminex assay has become the most widely used in clinical laboratories due to both increased sensitivity and specificity versus enzyme-linked immunosorbent assay. However, even the Luminex technique has shown some critical issues, and choosing the most reliable method still remains challenging. In this study, we verified the concordance of the results obtained in detecting anti-HLA antibodies with 2 kit vendors that provide reagents for the Luminex platform. Materials and Methods: We used 314 serum samples from patients on wait lists for solid-organ transplant. Sera were tested with LABScreen Mixed-LSM12 (One Lambda-Thermo Fisher, Canoga Park, CA, USA) and LIFECODES LifeScreen Deluxe-LMX (Gen-ProbeImmucor, Stanford, CT, USA),which we indicated as vendor A and vendor B, respectively. Anti-HLA class I and class II antibody analyses were conducted by verifying the concordance of the results with Cohen kappa coefficient statistics and confidence interval. Results:The kappa coefficient statistics showed "substantial" reliability for class I (0.61; confidence interval, 0.50-0.73) and "moderate" reliability for class II (0.56; confidence interval, 0.43-0.69). There were no considerable differences in results between the 2 kits regarding overall assignment of negativity or positivity of a sample. Discordant data between positive values for a test and negative for the other were found for samples with weak antibody positivity. Conclusions: Some discordant data were probably attributable to several factors such as the composition of the kits, the antibody titer in the serum, whether sera were diluted, different washing methods, and type of plate used.

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“…In contrast, the donor/recipient XMT as well as the anti-HLA antibody detection among the recipients play very important roles in the prevention of allograft rejection ESRD (5,17,18). In this concern, the overall rate of donor/recipient CDC-XMT and/or anti-HLA antibody positivity found in our patients of around 26% is significantly lower than that reported in the USA, where such positivity has been found to be as high as 40% (19).…”

Section: Discussionmentioning

confidence: 99%