BACKGROUND: Rapid detection of pathogens in blood is important in patient management, because the mortality rate associated with bloodstream infections is very high. We evaluated the efficiency of a 16S rDNA PCR assay for the detection of various pathogens in blood culture broth in METHODS: 16S rDNA PCR was performed on 221 blood culture bottles consisting of 99 culturepositive and 122 culture-negative samples. The results were compared with conventional culture methods. We also compared the efficiency of three DNA extraction and purification methods using proteinase K, triton X-100, and benzyl alcohol-guanidine DNA extraction of blood culture broths. RESULTS: The 16S rDNA PCR method detected 95 (12 Staphylococcus aureus, 27 coagulase negative staphylococci, 10 enterococci, 5 streptococci, 37 gram negative bacilli, 4 corynebacteria) of 99 positive culture bottles. Four false-negative results were obtained for bottles containing 2 Corynebacterium, 1 Escherichia coli, and 1 S. aureus species. All 122 bottles that showed no blood culture growth were negative by 16S rDNA PCR. Overall, the sensitivity, specificity, positive predictive values and negative predictive values of 16S rDNA PCR relative to the culture results were 96.0%, 100%, 100%, and 96.8%, respectively. Among the three DNA extraction methods, the benzyl alcohol-guanidine method was most effective. CONCLUSION: The 16S rDNA PCR assay is a rapid and efficient means of detecting various pathogens in the blood and has great potential for use in the clinical microbiology laboratory.
Coagulase ; Corynebacterium ; DNA ; DNA, Ribosomal* ; Endopeptidase K ; Escherichia coli ; Humans ; Mortality ; Octoxynol ; Polymerase Chain Reaction* ; Sensitivity and Specificity ; Sepsis ; Staphylococcus aureus

BACKGROUND: Blood culture is an important procedure for the determination of the etiologic agent of septicemia. Analysis of the blood culture results can provide clinicians with very important information for the empirical treatment of patients. METHODS: In this study the blood cuture results at Chosun University Hospital during the years 2002 to 2005 were analysed to determine the species and antimicrobial susceptibility of the isolates. Blood culture bottles were incubated in BACTEC 9240 blood culture system; the isolates were identified by Vitek II, and antimicrobial susceptibility was tested by Vitek II system or the NCCLS disk diffusion method. RESULTS: Positive blood cultures were obtained from 1,520 (18.5%) patients. Among the microorganisms isolated from blood culture, 97.0% were aerobic and facultative anaerobic bacteria and 2.8% were fungi. Frequently isolated organisms in decreasing order were coagulase-negative staphylococci (CNS), Escherichia coli, Staphylococus aureus, Stenotrophomonas maltophilia, Serratia marcescens, and Klebsiella pneumoniae. The proportion of Pseudomonas aeruginosa isolates resistant to ceftazidime and imipenem was increased during the study period. CONCLUSION: E. coli was the most frequent etiologic agent of bacteremia except CNS, common contaminants of skin, at Chosun University Hospital. It seems to be necessary to enhance infection control measures to cope with an increasing number of the resistant bacteria to various antibiotics.
Anti-Bacterial Agents ; Bacteremia ; Bacteria ; Bacteria, Anaerobic ; Ceftazidime ; Diffusion ; Escherichia coli ; Fungi ; Humans ; Imipenem ; Infection Control ; Klebsiella pneumoniae ; Pseudomonas aeruginosa ; Sepsis ; Serratia marcescens ; Skin ; Stenotrophomonas maltophilia

BACKGROUND: While broth based antimicrobial susceptibility test methods work well for the detection of the majority of antimicrobial resistance mechanisms, antimicrobial resistance mechanism in some microorganisms may not be detected by these methods. The purpose of this study was to compare Vitek II system with a standard method for the ability to detect inducible clindamycin resistance in Staphylococcus aureus. METHODS: Of 200 clinical isolates of S. aureus tested, 183 were methicillin resistant (MRSA) and 17 were methicillin susceptible (MSSA). A disk approximation test (Clinical Laboratory Standards Institute; CLSI, Wayne, PA, USA) was performed as the standard method by placing standard erythromycin and clindamycin disks in adjacent positions. Vitek II ID-GPI (bioMerieux, Durham, NC, USA) was used for identification and Vitek AST-P536 (bioMerieux, Durham, NC, USA) for antimicrobial susceptibility tests. RESULTS: Clindamycin resistance rates of S. aureus tested by disk diffusion and Vitek II system were 89% and 56%, respectively. All but one inducible clindamycin resistant MRSA isolates were susceptible to clindamycin by Vitek II system. Five inducible clindmycin resistant MSSA isolates were all susceptible to clindamycin by Vitek II system. Vitek II system did not detect the inducible clindamycin resistance in S. aureus. CONCLUSIONS: Our results showed that Vitek II system was unacceptable for the detection of inducible clindamycin resistance in S. aureus. We suggests that the disk approximation test should be used to detect the inducible clindamycin resistance in S. aureus.
Clindamycin* ; Diffusion ; Erythromycin ; Methicillin ; Methicillin Resistance ; Methicillin-Resistant Staphylococcus aureus ; Staphylococcus aureus

BACKGROUND: In hepatocellular carcinoma (HCC), the frequency of p53 mutation and the association with hepatitis B virus (HBV) infection varies with geographic locations and risk factors. The aim of this study was to determine the frequency of codon 249 mutation of p53, p53 overexpression, and HBV DNA positivity and to observe the relationship between them in Korean HCC. METHODS: We analyzed overexpression of p53 in hepatoma tissue from 17 HCC patients by immunohistochemistry (IHC), specific mutations at the third base position of codon 249 by PCR-restriction fragment length polymorphism (PCR-RFLP) method, and presence of HBV by nested PCR. RESULTS: Although a point mutation at codon 250 was seen in one (5.8%) of 17 patients, no codon 249 mutations were found in the patient cohort. The p53 protein was overexpressed in 4 (23.5%) of 17 HCCs. PCR for HBV DNA from HCCs showed a positivity rate of 82.4% (14 of 17 specimens). CONCLUSION: In HCC of this study, HBV infection was not associated with either 249 mutation or overexpression of p53, and overexpression of p53 protein seemed to be related to other than this mutation.
Carcinoma, Hepatocellular* ; Codon* ; Cohort Studies ; DNA ; Geographic Locations ; Hepatitis B virus* ; Hepatitis B* ; Hepatitis* ; Humans ; Immunohistochemistry ; Point Mutation ; Polymerase Chain Reaction ; Risk Factors

BACKGROUND: Group A rotavirus is a major cause of severe diarrhea in children throughout the world. For the proper management of rotavirus infections, it will be helpful to know their clinical characteristics according to the G and P genotypes of the infecting virus. METHODS: The diarrheal stool specimens from patients hospitalized in Chosun University Hospital during 2002-2003 were tested for rotavirus by Dipstick 'Eiken' Rota kit. Rotavirus antigen-positive stool specimens were analyzed for group A rotavirus by RT-PCR, and the group A-positive PCR products were genotyped for P and G types by PCR. RESULTS: Among the 119 specimens analyzed for genotypes, the predominant strain was genotype G4P[6] (51.3%), followed by G2P[4] (19.3%), G1P[8] (7.6%), G3P[8] (5.0%), and G9P[8] (4.2%). To examine the characteristics of each rotavirus genotype, a clinico-epidemiological study was performed for 100 patients whose medical records were available. The frequencies of diarrhea, vomiting, dehydration, and fever; the rates of nosocomial infection and transfer from other hospitals; and the mean severity scores were significantly different among the patients infected with different types of rotavirus. Especially, patients with G4P[6] type were more likely than those infected with other genotypes to show the following distinct features: Most patients showed milder symptoms and were neonates transferred from other obstetric hospitals and 68.4% of the cases were nosocomial infection. G4P[6] strains were isolated almost all along the year. The mean severity scores of patients infected by G4P[6], G2P[4], G1P[8], G3P[8], and G9P[8] strains were 6.8, 9.5, 8.0, 9.0, and 10.8, respectively. CONCLUSIONS: Many features of rotavirus infections including the epidemic period, rate of nosocomial infection, age and severity of symptoms were different according to the genotypes of the infecting virus.
Child ; Cross Infection ; Dehydration ; Diarrhea ; Fever ; Gastroenteritis* ; Genotype* ; Humans ; Infant, Newborn ; Medical Records ; Polymerase Chain Reaction ; Rotavirus Infections ; Rotavirus* ; Vomiting