However, the relationship between band intensity and final confirmation was not uniform among the onconeural antibodies tested

However, the relationship between band intensity and final confirmation was not uniform among the onconeural antibodies tested. was highly variable among the antibodies tested, from 7.2% (PNS+2 blot) and 5.8% (EUROLINE) for anti-Yo to 88.2% (PNS+2 blot) and 65.0% (EUROLINE) for anti-Hu. None of the 27 fragile positive sera by EUROLINE was confirmed. Band intensity in confirmed cases was variable among the antibodies from strong positive for those anti-Yo (n = 3) and anti-Hu (n = 11) to positive (n = 19) or strong positive (n = 9) for anti-SOX1. Among individuals having a nonconfirmed EUROLINE effect and available medical information, all experienced an alternative analysis, and only 6.7% had malignancy. Conclusions Immunodots may be useful for PNS screening, but a threshold should be established for each antibody, and medical info and confirmation by additional techniques are essential. Classification of evidence The study Panulisib (P7170, AK151761) provides Class IV evidence that immunodot assays for onconeural antibodies accurately determine individuals with paraneoplastic neurologic syndromes. Paraneoplastic neurologic syndromes (PNSs) are rare but now well-characterized immune-mediated neurologic diseases triggered by malignancy and diagnosed Panulisib (P7170, AK151761) by the presence of circulating autoantibodies.1 Among them, autoantibodies directed against intracellular neural antigens (also known as onconeural antibodies) are strongly associated with the presence of Panulisib (P7170, AK151761) an underlying cancer, and its detection is a cornerstone of PNS analysis. Indirect immunofluorescence (IIF) on rat mind slices is the desired screening test for recognition of onconeural antibodies, but the result should be confirmed by a second technique, either Western blot or for some cases such as anti-delta/notch-like epidermal growth factorCrelated receptor (anti-Tr/DNER) by cell-based assays (CBAs).2,3 These techniques have been developed mainly in research laboratories and are not available for routine analysis. However, 2 commercial immunodot assays are currently promoted: PNS+2 blot (Ravo Diagnostika, Freiburg, Germany) and EUROLINE PNS 12 Ag (Euroimmun, Lbeck, Germany). These immunodot assays present the advantage to be very easily and quickly performed as they are fully automated; they also display several antibodies at the same time. However, very little is known about the reliability of these immunodot assays, as only a few published studies have analyzed the level of sensitivity for the detection of anti-CV2/CRMP5 (collapsin response-mediator protein-5) antibodies,4 and the level of sensitivity and specificity for anti-Ma2 antibodies,5 and anti-SOX1 antibodies.6 In our laboratory, we use commercial immunodot assays as the first step of biological PNS analysis for those onconeural antibodies. Herein, we analyzed the diagnostic yield of 2 commercial immunodots by investigating the proportion of positive results confirmed by alternative techniques, taking also into account the medical info when it was available. Methods This study is definitely a single-center PDK1 retrospective analysis of samples (sera) from individuals with suspicion of PNS that were analyzed in the French Research Center on Paraneoplastic Neurological Syndromes (Lyon, France). First, sera were screened by commercial immunodot assays, using PNS+2 blot (Ravo Diagnostika), from January 2016 to May 2017, and EUROLINE PNS 12 Ag (Euroimmun), from July 2017 Panulisib (P7170, AK151761) to November 2018. Only the sera that were found positive from the immunodot assay for at least one of the onconeural antibodies were further analyzed by 2 in-house techniques: IIF followed by a technique using recombinant protein, either a Western blot for anti-CV2/CRMP5 and anti-amphiphysin antibodies or a CBA for the additional antibodies. When a positive immunodot result was also found positive using the 2 2 different confirmatory techniques (IIF and Western blot/CBA), the case was considered as confirmed. When both IIF and the third technique were bad, the immunodot result was considered as nonconfirmed. All confirmed cases were included in the database of the French Research Center, along with medical information. For the current study, we also collected medical data (including medical phenotype, malignancy association, and final analysis) for individuals whose serum was tested using the EUROLINE Panulisib (P7170, AK151761) PNS 12 Ag (Euroimmun) but were nonconfirmed; these data were not available for.