This study was purposed to investigate the effectiveness of bacterial screening with 24 hours holding in preventing and controlling bacterial contamination of platelets. Bacterial screening of apheresis platelets preserved for 24 hours was performed by using BacT/ALERT automatic bacterial culture system. The samples from 5 bags of platelet were taken in aseptic condition and were merged into 1 bag. The final sample was inoculated into aerobic and anaerobic bottle respectively for testing, meanwhile the screened platelet samples were held for 24 hours. If the platelets were cultured for 24 hours and identification of bacterial strains showed negative, the platelets could be released, and the original platelet samples should be rescreened if initiate positive was found. The results showed that in screening 8017 samples of apheresis platelets the initiate positive results were 16 (0.2%) and confirmed positive were 4 (0.05%). Out of 4 confirmed positive strains, three were Staphylococcus aureus and another was Staphylococcus auricularis. It is concluded that it is necessary for blood center to apply the method of bacterial screening of platelet with 24 hours holding as conventional screening method, which is an effective and feasible way to prevent and control bacterial contamination of platelets.
Bacteria ; isolation & purification ; Bacterial Infections ; prevention & control ; Bacteriological Techniques ; instrumentation ; Blood Platelets ; microbiology ; Blood Preservation ; methods ; standards ; Humans ; Platelet Transfusion ; adverse effects ; Plateletpheresis ; instrumentation

BACKGROUND: This study was designed as a quasi-experiment to evaluate automatic inoculation of fecal specimens, using the automated specimen inoculator Previ Isola (bioMerieux, France). METHODS: We evaluated the quality of cultures, recovery rates of enteropathogenic bacteria (Salmonella, Shigella, Campylobacter, and Yersinia species), and cost-effectiveness in terms of technical time. The Previ Isola recovery rates for the two-year period from August 2009 to July 2011 were compared with historical manual inoculation data of the previous two years (August 2007 to July 2009). The regional (Baden-Wurttemberg) and nationwide (Germany) trends of recovery rates for this four-year period were referred. RESULTS: A total of 5,884 fecal specimens were collected over the study period. Most positive cultures were for Salmonella, followed by Campylobacter. Compared with the historical data, the numbers of Campylobacter-positive specimens for a year between August and July were increased significantly, from 19 in 2007-2008 and 10 in 2008-2009 to 32 in 2009-2010 (P=0.002) and 32 in 2010-2011 (P=0.003), respectively. During the study period, the official data for our region and nationwide did not show this increase in the recovery rate of Campylobacter. For Salmonella, Shigella, and Yersinia, no significant changes were observed. Compared with manual inoculation, the mean hands-on time with Previ Isola inoculation was significantly shortened, from 37:30 min to 8:42 min per 15 fecal specimens. CONCLUSIONS: Inoculation by Previ Isola improves the quality of routine culture of fecal specimens, with better sensitivity for Campylobacter and less hands-on time.
Automation ; Bacteria/*isolation & purification ; Bacteriological Techniques/*methods/standards ; Campylobacter/isolation & purification ; Feces/*microbiology ; Humans ; Quality Control ; Salmonella/isolation & purification ; Shigella/isolation & purification ; Yersinia/isolation & purification

A multiplex PCR method was developed to detect the main pathogens of Qinghai Tibetan sheep endometritis. First, the genomes of five standard bacterial strains were extracted and specific primers were selected; the multiplex PCR method was established by using the genome of the standard strain as a template. The samples were collected by sterile cotton swab from Tibetan sheep uterus, and then placed in LB medium and numbered. After 48 h, the genomes of cultured bacteria were extracted and detected by single PCR method, then the positive samples were recorded. The positive samples detected by single PCR were selected for multiplex PCR detection and recorded again. The coincidence rate between these two methods was calculated to measure the accuracy of multiplex PCR. In order to identify the species of the pathogen, 30 positive samples verified by single and multiplex PCR were randomly selected for bacterial isolation and identification. In the 600 samples, the infected ratio of Streptococcus agalactiae (GBS) was 47.33%, Escherichia coli 34.83%, Staphylococcus aureus 6.5%, Salmonella and Trueperella pyogenes were negatively detected. Among the positive samples detected by multiplex PCR, the positive ratio of GBS was 45.50%, E. coli 33.50%, S. aureus 6.5%. Comparison of two detection results, Multiplex PCR detection coincidence rate is more than 95%. The isolated pathogens were identified as E. coli, GBS and S. aureus, which was consistent with the results of two methods. The multiplex PCR method was successfully established and the main pathogens of endometritis in Qinghai Tibetan sheep were GBS, E. coli and S. aureus.
Animals ; Bacteria ; genetics ; isolation & purification ; Bacteriological Techniques ; methods ; Endometritis ; microbiology ; veterinary ; Female ; Multiplex Polymerase Chain Reaction ; standards ; Polymerase Chain Reaction ; veterinary ; Sensitivity and Specificity ; Sheep ; Sheep Diseases ; microbiology ; Tibet

PURPOSE: Diagnosis of extrapulmonary tuberculosis (EPTB) poses serious challenges. A careful selection of appropriate gene targets is essential for designing a multiplex-polymerase chain reaction (M-PCR) assay. MATERIALS AND METHODS: We compared several gene targets of Mycobacterium tuberculosis, including IS6110, devR, and genes encoding MPB-64 (mpb64), 38kDa (pstS1), 65kDa (hsp65), 30kDa (fbpB), ESAT-6 (esat6), and CFP-10 (cfp10) proteins, using PCR assays on 105 EPTB specimens. From these data, we chose the two best gene targets to design an M-PCR. RESULTS: Among all gene targets tested, mpb64 showed the highest sensitivity (84% in confirmed cases and 77.5% in clinically suspected cases), followed by IS6110, hsp65, 38kDa, 30kDa, esat6, cfp10, and devR. We used mpb64+IS6110 for designing an M-PCR assay. Our M-PCR assay demonstrated a high sensitivity of 96% in confirmed EPTB cases and 88.75% in clinically suspected EPTB cases with a high specificity of 100%, taking clinical diagnosis as the gold standard. CONCLUSION: These M-PCR results along with the clinical findings may facilitate an early diagnosis of EPTB patients and clinical management of disease.
Bacteriological Techniques/methods ; DNA Transposable Elements/genetics ; DNA, Bacterial/analysis/genetics ; Early Diagnosis ; Female ; Gene Amplification ; Humans ; Male ; Multiplex Polymerase Chain Reaction/*methods ; Mycobacterium tuberculosis/genetics/*isolation & purification ; Polymerase Chain Reaction/*methods/standards ; Sensitivity and Specificity ; Tuberculosis/*diagnosis