(XLSX) Click here for extra data document.(11K, xlsx) Acknowledgments Component of the ongoing function was presented on the 27th Western european Congress of Clinical Microbiology and Infectious Illnesses, which occurred in Vienna, Austria, april 2017 from 22C25. inside the paper and its own Supporting Information data files. Abstract Cat damage disease (CSD) can be an infectious disease due to indirect immunofluorescence assays (IFAs), real-time polymerase string response (PCR)-structured recognition of is now being proposed as a more sensitive tool to diagnose CSD. Thus, here we have assessed the efficacy of real-time PCR in detecting in different specimens from patients with suspected CSD and compared it to that of IFA. From March 2011 to May 2016, at the Microbiology and Virology Unit, Azienda Ospedaliera Universitaria Citt della Salute e della Scienza di Torino, Turin, Italy, 115 clinical specimens (56 aspirated pus, 39 fresh lymph node AWD 131-138 biopsies, and 20 whole blood samples) and 99 sera from 115 patients with suspected CSD (62 females and 53 males between the ages of 3 months and 68 years) were analyzed by both real-time PCR, used in a qualitative way, and IFA (IgM and IgG) AWD 131-138 for the presence of DNA positivity was detected by real-time PCR in 37.39% of patients, while 62.61% of them were negative. Thus, patients were divided into two groups: real-time PCR+ (n = 43) and real-time PCR- (n = AWD 131-138 72). Real-time PCR screening of whole blood, biopsies, and aspirated pus revealed positivity in 40%, 38.46%, and 35.71% of patients, respectively. When we analyzed samples by IFA, we found the presence of in 28 out of 99 (28.28%) patients, of which 11 (11.11%) belonged to the real-time PCR+ group and 17 (17.17%) to the real-time PCR- group. Among the 71 seronegative subjects, 16 (16.16%) were found positive for by real-time PCR. Thus, by combining the results of both assays, we were able to increase the percentage AWD 131-138 of positive specimens from 27.27% (real-time PCR) or 28.28% (IFA) to 44.44% (real-time PCR+IFA). Altogether, these findings indicate that the early detection of in patients with suspicious CSD through combined real-time PCR and serological analyses can lead to a more accurate diagnosis of CSD, thereby allowing prompt and appropriate disease management. Introduction Cat scratch disease (CSD) is an emerging infectious disease worldwide caused pHZ-1 by from patient specimens [1,15], serology has later become the first-line diagnostic test for CSD, which is normally carried out by means of commercially available indirect immunofluorescence assays (IFAs) able to detect IgM and IgG antibodies to [11,16]. However, IFAs have low specificity and sensitivity, with results varying across laboratories due to between-kit variability [15,17,18]. Real-time polymerase chain reaction (PCR) on lymph nodes or other clinical samples has been more recently proposed as a suitable method to detect DNA in suspected cases of CSD due to its high sensitivity and specificity [12,19,20]. However, this technique is however limited by the requirement of invasive sampling such as lymphadenectomy or biopsy [11], which may be overcome by performing real-time PCR on DNA samples from aspirated pus or blood [17,21]. Indeed, real-time PCR has been successfully employed by two laboratories to detect DNA from blood of immunocompetent CSD patients, although this method may not be indicated in patients without bacterial DNAemia [17,21]. In this study, we have assessed the efficacy of real-time PCR IFA in detecting in a population-based cohort of patients with clinical presentations consistent with CSD. Our results suggest that a combined molecular and serological approach may improve the diagnosis of CSD. Materials and methods Ethics statement The ethical committee approval for the present research was not required as the patient samples (i.e. blood, aspirated pus, biopsy) were routinely subjected to microbiological evaluation at the Azienda Ospedaliero Universitaria (AOU) Citt della Salute e della Scienza di Torino, Turin, Italy. Informed written consent was obtained from all patients and from parents or guardians of the minors included in the study. The study was conducted in accordance with ethical standards and the Helsinki Declaration. Furthermore, to guarantee patient privacy, specimens were processed anonymously, and clinical data were blindly analyzed. All clinical specimens were coded to conceal patients identity and diagnosis. Patient population From March 2011 to May 2016, 115 clinical specimens (56 aspirated pus, 39 fresh lymph node biopsies, and 20 whole.