Human enteric Adenovirus-41 (HuEAdV-41) causes gastroenteritis, which detected by the polymerase chain reaction (PCR) base diagnostic system for clinical, food, environmental, fish and shellfish samples. We developed improved PCR and nested PCR primer set which had high specificity, sensitivity and reduced times. In this study, we compared seventeen conditions reported in the previous study that was using the PCR based HuEAdV-41 detection system, and non-enteric Adenovirus were detected in nine conditions. The most sensitive detection condition was up to 25 copies however it took 184 minutes of PCR reaction time. In this study, the PCR primer set developed had same level of sensitivity, it reduced the time of detection for clinical, food and seafood samples to 112 minutes. Developed nested PCR primer set needed 112 minutes but detected up to approximately 1 copy. In addition, developed PCR and nested PCR primer set was validated with twenty samples of underground water at random, of which ten samples showed specific band without non-specific reaction. We expect this study will be used to diagnose HuEAdV-41 from various samples.

This study was performed to assess microbial contamination of Aronia melanocarpa, blueberry, raspberry, and cranberry sold in several markets. We investigated total aerobic bacteria and detected foodborne bacteria by multiplex PCR from Aronia melanocarpa, blueberry, raspberry, and cranberry. Total aerobic bacteria of each sample showed mean 3.54 log CFU/g for Aronia melanocarpa, mean 1.90 log CFU/g for blueberry, and mean 1.40 log CFU/g for raspberry, but not detected in cranberry. Specially, Aronia melanocarpa contained high total aerobic bacteria contamination among various berries and contamination level reached 4.17 log CFU/g in sample 5. To evaluate the effect of distribution conditions, we also investigated total aerobic bacteria of various berries. Total aerobic bacteria showed mean 2.89 log CFU/g for berries in refrigerated distribution and 1.40 log CFU/g in frozen distribution, but not in dry distribution. For assessment of foodborne bacteria contamination, we conducted PCR with multiplex primers of E. coli O157, S. aureus, B. cereus, V. parahaemolyticus, L. monocytogenes, Y. enterocolitica, Salmonella spp., Shigella spp. Among these foodborne bacteria, B. cereus was amplified in Aronia melanocarpa in sample 4 and blueberry in sample 1, 2, 3, and 5. The result of quantitative analysis of B. cereus contamination showed 4.08 log CFU/g of Aronia melanocarpa in sample 4 and higher contamination rate 4.07 log CFU/g of blueberry in sample 3. These results suggest that strict food safety control in harvest and distribution of various berries is necessary to prevent foodborne disease and improve microbiological safety.

For the specific detection of human Parvovirus B19 (HuPaV-B19), we designed ten specific PCR primers from 3,800~4,500 nucleotides of HuPaV-B19 complete genome (NC_000883.2). Seventeen candidate PCR primer sets for specific detecting HuPaV-B19 were constructed. In specific reaction of HuPaV-B19, seventeen PCR primer sets showed specific band, however five PCR primer sets were selected basis of band intensity, amplicon size and location. In nonspecific reaction with seven reference viruses, four PCR primer sets showed non-specific band, however one PCR primer set is not. Detection sensitivity of final selective PCR primer set was 100 fg/μL for 112 minute, and PCR amplicon is 539 base pairs (bp). In addition, nested PCR primer set was developed, for detection HuPaV-B19 from a low concentration of contaminated samples. Selection of nested PCR primer set was basis of sensitivity and groundwater sample tests. Detection sensitivity of final selective PCR and nested PCR primer sets for the detection of HuPaV-B19 were 100 fg/μL and 100 ag/ μL basis of HuPaV-B19 plasmid, it was able to rapid and highly sensitive detection of HuPaV-B19 than previous reports. We expect developed PCR primer set in this study will used for detection of HuPaV-B19 in various samples.