ABSTRACT
Abstract Detection of Salmonella is very important to minimize the food safety risk. In this study, the recombinant PagC protein and PagC antibody were prepared and coupled with immunomagnetic beads (IMBs) to capture Salmonella cells from pork and milk samples. And then the SYBR Green qualitative PCR was developed to detect the pathogenic Salmonella. The results showed that the PagC polyclonal antiserum is of good specificity and the capture rate of 0.1 mg IMBs for Salmonella tended to be stable at the range of 70-74% corresponding to the concentrations between 101 and 104 CFU/mL. The method developed demonstrated high specificity for the positive Salmonella samples when compared to non-specific DNA samples, such as Escherichia coli, Staphylococcus aureus, Yersinia enterocolitica, and Yersinia pseudotuberculosis. The limit of detection of this assay was 18 CFU/mL. Detection and quantitative enumeration of Salmonella in samples of pork or milk shows good recoveries of 54.34% and 52.07%. In conclusion, the polyclonal antibody of recombinant PagC protein is effective to capture Salmonella from detected samples. The developed pagC antibody IMBs-qPCR method showed efficiency, sensitivity and specificity for 30 Salmonella detection, enabling detection within 10 h, which is a promising rapid method to detect Salmonella in emergency.
ABSTRACT
Abstract Detection of Salmonella is very important to minimize the food safety risk. In this study, the recombinant PagC protein and PagC antibody were prepared and coupled with immunomagnetic beads (IMBs) to capture Salmonella cells from pork and milk samples. And then the SYBR Green qualitative PCR was developed to detect the pathogenic Salmonella. The results showed that the PagC polyclonal antiserum is of good specificity and the capture rate of 0.1 mg IMBs for Salmonella tended to be stable at the range of 70-74% corresponding to the concentrations between 101 and 104 CFU/mL. The method developed demonstrated high specificity for the positive Salmonella samples when compared to non-specific DNA samples, such as Escherichia coli, Staphylococcus aureus, Yersinia enterocolitica, and Yersinia pseudotuberculosis. The limit of detection of this assay was 18 CFU/mL. Detection and quantitative enumeration of Salmonella in samples of pork or milk shows good recoveries of 54.34% and 52.07%. In conclusion, the polyclonal antibody of recombinant PagC protein is effective to capture Salmonella from detected samples. The developed pagC antibody IMBs-qPCR method showed efficiency, sensitivity and specificity for 30 Salmonella detection, enabling detection within 10 h, which is a promising rapid method to detect Salmonella in emergency.
ABSTRACT
Abstract Detection of Salmonella is very important to minimize the food safety risk. In this study, the recombinant PagC protein and PagC antibody were prepared and coupled with immunomagnetic beads (IMBs) to capture Salmonella cells from pork and milk samples. And then the SYBR Green qualitative PCR was developed to detect the pathogenic Salmonella. The results showed that the PagC polyclonal antiserum is of good specificity and the capture rate of 0.1 mg IMBs for Salmonella tended to be stable at the range of 70-74% corresponding to the concentrations between 101 and 104 CFU/mL. The method developed demonstrated high specificity for the positive Salmonella samples when compared to non-specific DNA samples, such as Escherichia coli, Staphylococcus aureus, Yersinia enterocolitica, and Yersinia pseudotuberculosis. The limit of detection of this assay was 18 CFU/mL. Detection and quantitative enumeration of Salmonella in samples of pork or milk shows good recoveries of 54.34% and 52.07%. In conclusion, the polyclonal antibody of recombinant PagC protein is effective to capture Salmonella from detected samples. The developed pagC antibody IMBs-qPCR method showed efficiency, sensitivity and specificity for 30 Salmonella detection, enabling detection within 10 h, which is a promising rapid method to detect Salmonella in emergency.
Subject(s)
Animals , Salmonella/isolation & purification , Food Contamination , Immunomagnetic Separation/methods , Real-Time Polymerase Chain Reaction/methods , Food Microbiology/methods , Salmonella/genetics , Bacterial Proteins/immunology , Sensitivity and Specificity , Milk/microbiology , Meat/microbiology , Antibodies, Bacterial/immunology , Antibodies, Bacterial/metabolismABSTRACT
In order to explore the possibility to predict the risk factors for postoperative complications and survival time, the clinical data of 152 patients (including 116 males and 36 females) who had undergone neo-adjuvant therapy and surgery for stage Ilia and B non-small cell lung cancer (NSCLC) were retrospectively analyzed. Demographic data, preoperative functional parameters,staging, induction regimen (chemotherapy alone or associated with radiotherapy), associated disorders, and data about operation were collected. Chi-square test and multivariate analysis fitting the unconditional logistic regression model were performed to identify predictors of postoperative complications, while Kaplan-Meier and multivariate Cox proportional hazard model were employed to identify predictors of survival time, respectively. The univariate analysis demonstrated that forced expiratory volume in 1 second predicted percent (FEVI%, P=0.040) and associated disorders (P=0.020) were the predictive factors of complications, but multivariate analysis found no independence factors (P>0.05) of it. Univariate Kaplan-Meier analysis showed that stage (P=0.050) and pneumonectomy (P=0.018) affected the survival time. However, multivariate Cox proportional hazard model analysis demonstrated that only pneumonectomy (P=0.026) was associated with a decreased survival time, but no differences between right and left pneumonectomy were found. The results suggest that the risk factor for postoperative complications is acceptable, and pneumonectomy is associated with increased mortality, which should be performed only in stage Ⅲ NSCLC patients.