BIOTECON diagnostics foodproof Listeria monocytogenes Detection Kit, 5' nuclease in combination with the foodproof ShortPrep II Kit.

A method was developed for the detection of Listeria monocytogenes in food. The method is based on real-time PCR using hydrolysis probes (5' Nuclease). This advanced PCR method was designed to reduce the time necessary to achieve results from PCR reactions and to enable the user to monitor the amplification of the PCR product simultaneously, in real-time. After DNA isolation using the BIOTECON foodproof ShortPrep II Kit designed for the rapid preparation of L. monocytogenes DNA for direct use in PCR, the real-time detection of L. monocytogenes DNA is carried out using the foodproof Listeria monocytogenes Detection Kit. The kit provides primers and hydrolysis probes for sequence-specific detection, convenient premixed reagents, and controls for reliable interpretation of results. For the internal comparison study, three different foods (soft cheese, coalfish, and smoked ham) were analyzed, chosen from the 15 food groups recommended by the AOAC Research Institute for detection of L. monocytogenes. From each food, 20 samples were inoculated with a low level (1-10 CFU/25 g) and 20 samples with a high level (10-50 CFU/25 g) of L. monocytogenes. Additionally, five nonspiked samples were prepared from each food. Depending on the matrix, the food samples were examined with the test kits and compared with the cultural methods according to the U.S. Food and Drug Administration's Bacteriological Analytical Manual or the U.S. Department of Agriculture/Food Safety and Inspection Service Microbiology Laboratory Guidebook.

BIOTECON diagnostics foodproof E. coli O157 detection kit, 5' nuclease for E. coli O157 in combination with foodproof ShortPrep II Kit.

The method describes the detection of Escherichia coli O157 in food. The method is based on real-time PCR using hydrolysis probes (5' Nuclease). This advanced PCR method was designed to reduce the time necessary to achieve results from PCR reactions and enable the user to monitor the amplification of the PCR product simultaneously in real time. After DNA isolation using the BIOTECON foodproof ShortPrep II Kit designed for the rapid preparation of E. coli O157 DNA for direct use in PCR, the real-time detection of E. coli O157 DNA is carried out using the foodproof E. coli O157 Detection Kit. The kit provides primers and hydrolysis probes for sequence-specific detection, convenient premixed reagents, and controls for reliable interpretation of results. For repeatability studies three different foods (egg salad, large bockwurst/frankfurter, and apple juice) were analyzed, chosen from the 15 food groups recommended by the AOAC Research Institute for E. coli O157 detection. From each food, 20 samples were inoculated with a low level (1-10 CFU/25 g) and 20 samples with a high level (10-50 CFU/25 g) ofE. coli O157. Additionally, five nonspiked samples were prepared from each food. Depending on the matrix, the food samples were examined with the test kits and compared with the cultural methods according to the U.S. Food and Drug Administration Bacteriological Analytical Manual or the U.S. Department of Agriculture/Food Safety and Inspection Service Microbiology Laboratory Guidebook.

Promoting undergraduate interest, preparedness, and professional pursuit in the sciences: An outcomes evaluation of the SURE program at Emory University.

We report on an outcomes assessment of the Summer Undergraduate Research Experience (SURE) Program at Emory University in Atlanta, GA. Using follow-up survey data and academic transcripts, we gauge SURE's impact on levels of interest in, preparedness for, and actual pursuit of graduate study and professional careers in the sciences for the program's first 15 summer cohorts (1990-2004). Our follow-up survey indicated significant increases in all research preparedness skills considered, notably in ability to give a poster research presentation, to discuss research at a graduate school interview, and to apply research ethics principles. About a third of SURE graduates went on to complete a graduate degree >90% considered SURE as important or very important in their academic development. Respondents reported postprogram increases in the level of interest in academic and research careers, and reported high levels of employment in science careers and job satisfaction. Regression analyses of Emory SURE participant transcripts revealed that participants take significantly more science courses as seniors and earn higher grades in those courses than nonparticipants. This trend held after correcting for indicators of prior interest (first-year course work, GPA, and math SAT scores), gender, and minority status. We also report on an external survey completed by SURE participants.

Foodproof Salmonella Detection Kit. Performance Tested Method 120301.

The foodproof Salmonella Detection Kit was previously validated in the Performance Tested Methods program for the detection of Salmonella species in a variety of foods, including milk powder, egg powder, coconut, cocoa powder, chicken breast, minced meat, sliced sausage, sausage, smoked fish, pasta, white pepper, cumin, dough, wet pet food, dry pet food, ice cream, watermelon, sliced cabbage, food dye, and milk chocolate. The method was shown to be equivalent to the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA-BAM) and the U.S. Department of Agriculture, Food Safety and Inspection Service's Microbiology Laboratory Guidebook reference culture procedures. In the first Emergency Response Validation (ERV) extension study, peanut butter was inoculated with S. enterica. ser Typhimurium. For the low inoculation level (1.08 CFU/25 g), a Chi-square value of 2.25 indicated that there was no significant performance difference between the foodproof Salmonella Detection Kit and the FDA-BAM reference method. For high-level inoculation (11.5 CFU/25 g) and uninoculated control, there was 100% agreement between the methods. In the second ERV extension study, peanut butter was inoculated with S. enterica. ser Typhimurium. For both inoculation levels (0.1 and 0.5 CFU/25 g by most probable number), Chi-square values of 0 indicated that there was no significant performance difference between foodproof Salmonella Detection Kit and the FDA-BAM reference method.

Roche/BIOTECON Diagnostics LightCycler foodproof L. monocytogenes detection kit in combination with ShortPrep foodproof II Kit. Performance-Tested Method 070401.

A method was developed for the detection of L. monocytogenes in food based on real-time polymerase chain reaction (PCR). This advanced PCR method was designed to reduce the time needed to achieve results from PCR reactions and to enable the user to monitor the amplification of the PCR product simultaneously, in real-time. After DNA isolation using the Roche/BIOTECON Diagnostics ShortPrep foodproof II Kit (formerly called Listeria ShortPrep Kit) designed for the rapid preparation of L. monocytogenes DNA for direct use in PCR, the real-time detection of L. monocytogenes DNA is performed by using the Roche/BIOTECON Diagnostics LightCycler foodproof L. monocytogenes Detection Kit. This kit provides primers and hybridization probes for sequence-specific detection, convenient premixed reagents, and different controls for reliable interpretation of results. For repeatability studies, 20 different foods, covering the 15 food groups recommended from the AOAC Research Institute (AOAC RI) for L. monocytogenes detection were analyzed: raw meats, fresh produce/vegetables, processed meats, seafood, egg and egg products, dairy (cultured/noncultured), spices, dry foods, fruit/juices, uncooked pasta, nuts, confectionery, pet food, food dyes and colorings, and miscellaneous. From each food 20, samples were inoculated with a low level (1-10 colony-forming units (CFU)/25 g) and 20 samples with a high level (10-50 CFU/25 g) of L. monocytogenes. Additionally, 5 uninoculated samples were prepared from each food. The food samples were examined with the test kits and in correlation with the cultural methods according to U.S. Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM) or U.S. Department of Agriculture (USDA)/Food Safety and Inspection Service (FSIS) Microbiology Laboratory Guidebook. After 48 h of incubation, the PCR method in all cases showed equal or better results than the reference cultural FDA/BAM or USDA/FSIS methods. Fifteen out of 20 tested food types gave exactly the same amount of positive samples for both methods in both inoculation levels. For 5 out of 20 foodstuffs, the PCR method resulted in more positives than the reference method after 48 h of incubation. Following AOAC RI definition, these were false positives because they were not confirmed by the reference method (false-positive rate for low inoculated foodstuffs: 5.4%; for high inoculated foodstuffs: 7.1%). Without calculating these unconfirmed positives, the PCR method showed equal sensitivity results compared to the alternative method. With the unconfirmed PCR-positives included into the calculations, the alternative PCR method showed a higher sensitivity than the microbiological methods (low inoculation level: 100 vs 98.0%; sensitivity rate: 1; high inoculation level: 99.7 vs 97.7%; sensitivity rate, 1). All in-house and independently tested uninoculated food samples were negative for L. monocytogenes. The ruggedness testing of both ShortPrep foodproof II Kit and Roche/BIOTECON LightCycler foodproof L. monocytogenes Detection Kit showed no noteworthy influences to any variation of the parameters component concentration, apparatus comparison, tester comparison, and sample volumes. In total, 102 L. monocytogenes isolates (cultures and pure DNA) were tested and detected for the inclusivity study, including all isolates claimed by the AOAC RI. The exclusivity study included 60 non-L. monocytogenes bacteria. None of the tested isolates gave a false-positive result; specificity was 100%. Three different lots were tested in the lot-to-lot study. All 3 lots gave equal results. The stability study was subdivided into 3 parts: long-term study, stress test, and freeze-defrost test. Three lots were tested in 4 time intervals within a period of 13 months. They all gave comparable results for all test intervals. For the stress test, LightCycler L. monocytogenes detection mixes were stored at different temperatures and tested at different time points during 1 month. Stable results were produced at all storage temperatures. The freeze-defrost analysis showed no noteworthy aggravation of test results. The independent validation study examined by Campden and Chorleywood Food Research Association Group (CCFRA) demonstrated again that the LightCycler L. monocytogenes detection system shows a comparable sensitivity to reference methods. With both the LightCycler PCR and BAM methods, 19 out of 20 inoculated food samples were detected. The 24 h PCR results generated by the LightCycler system corresponded directly with the FDA/BAM culture results. However, the 48 h PCR results did not relate exactly to the FDA/BAM results, as one sample found to be positive by the 48 h PCR could not be culturally confirmed and another sample which was negative by the 48 h PCR was culturally positive.

Individual and structural influences shaping pharmacists' decisions to sell syringes to injection drug users in Atlanta, Georgia.

OBJECTIVE: To better understand the individual (e.g., attitudes and beliefs) and structural (e.g., laws and regulations) factors that influence and shape pharmacists' decisions about selling syringes to injection drug users (IDUs). DESIGN: Qualitative research. SETTING: Metropolitan Atlanta. PARICIPANTS: 20 practicing pharmacists who work in or near areas of high drug use in Atlanta, and nine pharmacists who are considered leaders in their profession in Georgia. INTERVENTIONS: Semistructured, in-depth interviews. MAIN OUTCOME MEASURES: Individual and structural factors that influence pharmacists' decisions about selling syringes to IDUs. RESULTS: Pharmacists reported that they use their professional discretion in making syringe sale decisions and that these decisions are influenced by individuals factors such as their personal attitudes and beliefs about the nature and causes of drug use, and by structural factors such as the Georgia Board of Pharmacy regulation stating that syringes cannot be sold if they will be used for an "unlawful purpose." CONCLUSIONS: IDUs' access to sterile syringes from pharmacies in Atlanta, would likely be increased by (1) providing practicing pharmacists with professional education programs that describe the broad professional support for IDU access to sterile syringes and why blood-borne infection prevention is a legitimate medical purpose for selling syringes and (2) removing or modifying the restrictive Board of Pharmacy regulation governing syringe sales.

Health services utilization by injection drug users participating in a needle exchange program.

OBJECTIVE: To determine the characteristics associated with health care and drug treatment utilization among a distinctly high-risk sub-population of injectors participating in a needle exchange program (NEP). METHODS: Between June 1998 and May 1999, study staff collected demographic and health services utilization data on participants of the Baltimore NEP. Odds ratios and logistic regression were used to identify the participant characteristics associateds with utilizing primary health care and drug treatment during the prior 3 years. RESULTS: Among 269 participants, 81% were African-American and 66% were male. Over half (56%) had not graduated from high school, 89% were unemployed, 70% did not have health insurance, and the median age was 39 years. Fifty-eight percent of the participants reported utilizing primary care (i.e., visited a physician or other health care provider) and 44% had utilized drug treatment during the prior 3 years. Primary care utilization was associated with age > or = 39 [adjusted odds ratio (AOR) = 1.82], having health insurance (AOR = 2.16), and exchanging a higher volume of syringes per NEP visit (AOR = 2.45). Recent drug treatment utilization was associated with African-American race (AOR = 0.41), unemployment (AOR = 2.72), having health insurance (AOR = 2.05), and exchanging a higher volume of syringes per NEP visit (AOR = 0.60). CONCLUSIONS: Health insurance was significantly associated with the recent utilization of both primary care and drug treatment, yet only one-third of NEP attenders were insured. Facilitating the uptake of health insurance services at NEP sites may improve the access to health care for drug users who are currently not utilizing the health care system.