• Korean Journal of Microbiology
• /
• v.47 no.2
• /
• pp.117-123
• /
• 2011

The purpose of this study was to evaluate and compare different elution and concentration methods for optimization of human rotavirus (HRV) detection method using real-time RT-PCR and cell culture techniques. The leafy vegetable samples (lettuce, Chinese cabbage) were artificially inoculated with HRV. Viruses were extracted from the vegetables by two different elution buffers, buffer A (100 mM Tris-HCl, 50 mM glycine, 3% beef extract, pH 9.5) and buffer B (250 mM Threonine, 300 mM NaCl, pH 9.5), and the extracted viruses were concentrated by filtration and PEG precipitation sequentially. To determine infectivity of the viruses, the viruses recovered from the samples were infected to the MA-104 cells, and integrated cell culture real-time RT-PCR was performed at 1, 48, 72, 96, 120, 144, 168 h post-infection (p.i.). The elution buffer A was more efficient in extracting the virus from the produce samples tested than the buffer B, 29.54% and 18.32% of recoveries, respectively. The sensitivity of real-time RT-PCR method was markedly improved when the virus was concentrated by the filtration method. When the viruses were eluted and concentrated by buffer A and filtration, respectively, the average recovery rate was approximately 51.89%. When the viruses recovered from samples were infected to MA-104 cell, infectious HRV was detected within 48 h p.i. by ICC/real-time RT-PCR, whereas cytopathic effects were not observed until 72 h p.i. The optimized detection method evaluated in this study could be useful for rapid and reliable detection of HRV in fresh produce products and applied for detection of other food-borne viruses.

• Pediatric Infection and Vaccine
• /
• v.13 no.1
• /
• pp.71-77
• /
• 2006

Purpose : Shigellosis is still an important cause of acute food-borne diarrheal diseases throughout the world. We investigated the transmission routes and clinical course through the outbreak of shigellosis in Mapo-Gu, Seoul. Methods : From October 7th to November 19th in 2005, Mapo-Gu public health center had referred 32 patients with confirmed or suspected shigellosis to the National Medical Center. We had investigated source of infection, epidemiology, laboratory findings, and clinical course of the cases occurred during this outbreak. Results : Among 32 patients, 24 patients had been confirmed with shigellosis, 8 patients had been diagnosed with suspected shigellosis. They ranged in age from 5 months to 12 years old and their mean age was 6.5 years. The clinical manifestations were as follows; diarrhea, fever, abdominal pain and asymptomatic condition. Symptoms had sustained for 3.7 days on the average. S. sonnei were cultured by rectal swab and founded to be resistant to ampicillin and TMP/SMX except to 3rd generation cephalosporin. After treatment with antibiotics such as cefixime and ceftriaxone or imipenem and conservative treatment with electrolyte and fluid replacement for 5~7 days, Stool cultures of the rectal swab grew no Shigella in these cases except 3 cases. Conclusion : An outbreak of shigellosis had occurred in a preschool and elementary school children. From the same results of antimicrobial susceptibility and pulsed-field gel electrophoresis patterns in this study, we suggest that the outbreak of shigellosis in this report had been originated from a single strain. According to all negative results about suspected food and water cultures, we couldn't find out source of infection. Through materials offerred by Mapo-Gu Public Health Center, we presumed the trasmission routes probably were person-to-person.

• Korean Journal of Environmental Agriculture
• /
• v.27 no.2
• /
• pp.156-162
• /
• 2008

Salmonellosis is a major bacterial zoonosis that causes a variety of disease syndromes, self-limited enteritis to fatal infection in animals and food-borne infection and typhoid fever in humans. Recently, the emergence of multidrug resistant strains of Salmonella spp. causes more serious problems in environment and public health. The present study was investigated the antibacterial effect of Houttuynia cordata ethanol extract(HCEE) for murine salmonellosis. In the cytotoxic effect of HCEE on RAW 264.7 cells, there was no detectable effect with any concentrations between 25 and 100 ${\mu}g/ml$ after 8 h incubation. The bacteriocidal effect of HCEE was not showed on a Salmonella enterica serovar Typhimurium(S. typhimurium). HCEE makes morphological change of the RAW 264.7 cells, and there was significant decreased bacterial uptake and intracellular replication within Salmonella infected cells. And further nitric oxide(NO) production of Salmonella infected RAW 264.7 cells with HCEE was decreased comparing to RAW 264.7 cells without HCEE until 8 h post infection. Oral administration of HCEE showed a therapeutic effect for S. typhimurium infected BALB/c mice. The mortality of HCEE treated mouse was 80% until 12 days, while that of HCEE untreated mouse was 100 % until 8 days after lethal dose of S. typhimurium infection. These data suggested that HCEE has a potency treatment for intracellular replicative pathogen including salmonellosis, brucellosis, tuberculosis, listeriosis etc., and the application of HCEE makes new strategies for safety medicine development without antibiotic resistance bacterial appearance and residue problem in food and solves the public health problem from antibiotic mis- and over use.