BACKGROUND: The determination of serotype of enteroviruses is useful for the discrimination between sporadic and epidemic infections. The conventional serotyping method is time-consuming and labor-intensive. Recently, molecular method was introduced for the serotyping of enteroviruses. The aim of this study was to establish a method to isolate and analyze enteroviruses from various specimens utilizing molecular biological techniques and to determine which strains were phylogenetically related to clinical samples. METHODS: Clinical samples in this study included 164 cerebrospinal fluid (CSF), 136 stool, 15 sera, 6 throat swab, 5 urine, and 4 sputa, which were obtained from hospitalized patients, primarily infants or children presenting symptoms of aseptic meningitis in 1998. RD cells were used for enterovirus isolation. RT-PCR was performed with RD cell lysate showing CPE. The primers 011 and 012 were used for the VP1 region, and the primers EN1 and EN2 for 5'-UTR. The nucleotide sequences of VP1 region were determined and analyzed with BLAST program. RESULTS: Among 333 samples, only 23 samples produced CPE: 17 samples at first and six samples at the second blind passage. Fifteen isolates were related to coxsackievirus B2 two to echovirus 4, three to echovirus 6, and three to echovirus 18. All 23 viral isolates displayed a nucleotide sequence identity of 80-95%, compared with the reference serotypes. However, the identity was increased up to 93-100% when the VP1 region was translated into amino acids CONCLUSIONS: Since CB2 type was 55% among enteroviral isolates, the CB2 was determined as the major causative serotype of enteroviral meningitis in 1998. CB2 type was emerged between June and July, EC4 and EC6 was limited to July, and EC18 was in August.
Amino Acids ; Base Sequence ; Cerebrospinal Fluid ; Child ; Discrimination (Psychology) ; Echovirus 6, Human ; Enterovirus B, Human ; Enterovirus* ; Humans ; Infant ; Meningitis* ; Meningitis, Aseptic ; Pharynx ; Serotyping*

BACKGROUND: The determination of serotype of enteroviruses is useful for the discrimination between sporadic and epidemic infections. The conventional serotyping method is time-consuming and labor-intensive. Recently, molecular method was introduced for the serotyping of enteroviruses. The aim of this study was to establish a method to isolate and analyze enteroviruses from various specimens utilizing molecular biological techniques and to determine which strains were phylogenetically related to clinical samples. METHODS: Clinical samples in this study included 164 cerebrospinal fluid (CSF), 136 stool, 15 sera, 6 throat swab, 5 urine, and 4 sputa, which were obtained from hospitalized patients, primarily infants or children presenting symptoms of aseptic meningitis in 1998. RD cells were used for enterovirus isolation. RT-PCR was performed with RD cell lysate showing CPE. The primers 011 and 012 were used for the VP1 region, and the primers EN1 and EN2 for 5'-UTR. The nucleotide sequences of VP1 region were determined and analyzed with BLAST program. RESULTS: Among 333 samples, only 23 samples produced CPE: 17 samples at first and six samples at the second blind passage. Fifteen isolates were related to coxsackievirus B2 two to echovirus 4, three to echovirus 6, and three to echovirus 18. All 23 viral isolates displayed a nucleotide sequence identity of 80-95%, compared with the reference serotypes. However, the identity was increased up to 93-100% when the VP1 region was translated into amino acids CONCLUSIONS: Since CB2 type was 55% among enteroviral isolates, the CB2 was determined as the major causative serotype of enteroviral meningitis in 1998. CB2 type was emerged between June and July, EC4 and EC6 was limited to July, and EC18 was in August.
Amino Acids ; Base Sequence ; Cerebrospinal Fluid ; Child ; Discrimination (Psychology) ; Echovirus 6, Human ; Enterovirus B, Human ; Enterovirus* ; Humans ; Infant ; Meningitis* ; Meningitis, Aseptic ; Pharynx ; Serotyping*

BACKGROUND: Shiga toxin-producing Escherichia coli (STEC) was found in several serotypes of E coli including 0157 serotype. Sorbitol-MacConkey agar: may be useful for the detection of E. coli 0157, but is not helpful for the detection of sorbitol-fermenting STEC other than 0157. Moreover, some strains of E. colt 0157 can ferment sorbitol. In this study, in situ hybridization using DNA probe of shiga toxin was used for the isolation of STEC from the PCR-positive stool and -Sequenbe analysis of a part of shiga toxin gene was performed. METHODS: The stool was incubated in LB broth overnight and DNA was extracted from the culture fluid. Multiplex PCR was performed with primers for stxl and stx2 genes. Specimen showed PCR-positive was incubated on MacConkey agar and colonies were blotted with nitrocellulose membrane. Digoxigenin-labelled DNA probe for shiga toxin was made by PCR and the positive colonies were detected with anti-digoxigenin-alkaline phosphatase conjugate and nitroblue tetrazolium. Agglutination test with antisera was performed for the serotying and VTEC-RPLA kit was used for the toxin production. Sequence analysis of PCR products was performed with automatic sequence analyser. RESULTS: An stxl-negative, but stx2-positive PCR was observed in a three-year-old girl, who visited Kumi Hospital on July 19, 1999 complaining of vomiting and diarrhea. The positive colonies were isolated by in situ hybridization using stx2-specific DNA probe. The titers of stxl and stx2 by VTEC-RPLA test were negative and 1:64, respectively. Agglutination for the serotyping was not observed with all of the 0 antisera. 160-nucleotide sequence of stx2 of this isolate was identical with bacteriophage 933W (GenBank X07865), except for the change (T-C) of 957th nucleotide and amino acid sequence was identical each other. CONCLUSIONS: For the sensitive detection of STEC from the stool of patients with diarrhea, multiplex PCR is recommended with stxl- and stx2-specific primers. And in situ hybridization should be performed in PCR-positive specimen for the isolation of STEC. This method may be helpful for the detection of STEC as the causative microorganisms in food-borne outbreak.
Agar ; Agglutination ; Agglutination Tests ; Amino Acid Sequence ; Bacteriophages ; Collodion ; Diarrhea ; DNA ; Escherichia coli ; Female ; Gyeongsangbuk-do ; Humans ; Immune Sera ; In Situ Hybridization ; Membranes ; Multiplex Polymerase Chain Reaction ; Nitroblue Tetrazolium ; Polymerase Chain Reaction ; Sequence Analysis* ; Serotyping ; Shiga Toxin ; Shiga-Toxigenic Escherichia coli* ; Sorbitol ; Vomiting

BACKGROUND: Shiga toxin-producing Escherichia coli (STEC) was found in several serotypes of E coli including 0157 serotype. Sorbitol-MacConkey agar: may be useful for the detection of E. coli 0157, but is not helpful for the detection of sorbitol-fermenting STEC other than 0157. Moreover, some strains of E. colt 0157 can ferment sorbitol. In this study, in situ hybridization using DNA probe of shiga toxin was used for the isolation of STEC from the PCR-positive stool and -Sequenbe analysis of a part of shiga toxin gene was performed. METHODS: The stool was incubated in LB broth overnight and DNA was extracted from the culture fluid. Multiplex PCR was performed with primers for stxl and stx2 genes. Specimen showed PCR-positive was incubated on MacConkey agar and colonies were blotted with nitrocellulose membrane. Digoxigenin-labelled DNA probe for shiga toxin was made by PCR and the positive colonies were detected with anti-digoxigenin-alkaline phosphatase conjugate and nitroblue tetrazolium. Agglutination test with antisera was performed for the serotying and VTEC-RPLA kit was used for the toxin production. Sequence analysis of PCR products was performed with automatic sequence analyser. RESULTS: An stxl-negative, but stx2-positive PCR was observed in a three-year-old girl, who visited Kumi Hospital on July 19, 1999 complaining of vomiting and diarrhea. The positive colonies were isolated by in situ hybridization using stx2-specific DNA probe. The titers of stxl and stx2 by VTEC-RPLA test were negative and 1:64, respectively. Agglutination for the serotyping was not observed with all of the 0 antisera. 160-nucleotide sequence of stx2 of this isolate was identical with bacteriophage 933W (GenBank X07865), except for the change (T-C) of 957th nucleotide and amino acid sequence was identical each other. CONCLUSIONS: For the sensitive detection of STEC from the stool of patients with diarrhea, multiplex PCR is recommended with stxl- and stx2-specific primers. And in situ hybridization should be performed in PCR-positive specimen for the isolation of STEC. This method may be helpful for the detection of STEC as the causative microorganisms in food-borne outbreak.
Agar ; Agglutination ; Agglutination Tests ; Amino Acid Sequence ; Bacteriophages ; Collodion ; Diarrhea ; DNA ; Escherichia coli ; Female ; Gyeongsangbuk-do ; Humans ; Immune Sera ; In Situ Hybridization ; Membranes ; Multiplex Polymerase Chain Reaction ; Nitroblue Tetrazolium ; Polymerase Chain Reaction ; Sequence Analysis* ; Serotyping ; Shiga Toxin ; Shiga-Toxigenic Escherichia coli* ; Sorbitol ; Vomiting

BACKGROUND: The polymerase chain reaction (PCR) test is important for the confirmatory diagnosis of tuberculosis (TB) caused by Mycobacterium tuberculosis. The aim of this study was to analyze the yield of repeated PCR testing in patients with confirmed pulmonary TB. METHODS: The medical records of 130 patients, who had more than two consecutive PCR tests and a M. tuberculosis-positive sputum culture from August, 2006 to December, 2007, were retrospectively reviewed for the purposes of this study. A positive TB-PCR test was defined as at least one positive test result. RESULTS: The cumulative positive PCR test rate was 80% (104/130), with gradually increasing rates of positive findings upon the first, second and third TB-PCR tests with 52.3%, 68.5% and 75.4%, respectively. However, further testing did not increase the positive rate further. CONCLUSION: Repeated PCR testing at least three times for M. tuberculosis is helpful for diagnosis of pulmonary TB.
Humans ; Medical Records ; Mycobacterium tuberculosis ; Polymerase Chain Reaction ; Retrospective Studies ; Sputum ; Tuberculosis ; Tuberculosis, Pulmonary

Rectus sheath hematoma (RSH) is a rare condition caused by hemorrhage into the rectus sheath. It is usually associated with severe cough, abdominal surgery, coagulopathy, and anticoagulation treatment. RSH can be difficult to diagnose and can be misdiagnosed as acute appendicitis, as diverticulitis, or as an ovarian mass. Although RSH usually presents as a benign condition, it can be life threatening, especially in the critically-ill patient. Here, we report a case of fatal RSH due to hypovolemic shock in a critically-ill 73-year-old woman, who had received heparin treatment due to acute myocardial infarction in the intensive care unit and who had been successfully treated by conservative management.
Aged ; Appendicitis ; Cough ; Diverticulitis ; Female ; Hematoma ; Hemorrhage ; Heparin ; Humans ; Hypovolemia ; Intensive Care Units ; Myocardial Infarction ; Rectus Abdominis ; Shock

The presence of epidermal growth factor receptor (EGFR) mutation is a prognostic and predictive marker for EGFR-tyrosine kinase inhibitor (TKI) therapy. However, inevitably, relapse occurs due to the development of acquired resistance, such as T790M mutation. We report a case of repeated responses to EGFR-TKIs in a never-smoked woman with adenocarcinoma. After six cycles of gemcitabine and cisplatin, the patient was treated by gefitinib for 4 months until progression. Following the six cycles of third-line pemetrexed, gefitinib retreatment was initiated and continued with a partial response for 6 months. After progression, she was recruited for an irreversible EGFR inhibitor trial, and the time to progression was 11 months. Although EGFR direct sequencing on the initial diagnostic specimen revealed a wild-type, we performed a rebiopsy from the progressed subcarinal node at the end of the trial. The result of peptide nucleic acid clamping showed L858R/L861Q.
Adenocarcinoma ; Cisplatin ; Constriction ; Deoxycytidine ; Epidermal Growth Factor ; Female ; Glutamates ; Guanine ; Humans ; Lung ; Lung Neoplasms ; Phosphotransferases ; Quinazolines ; Receptor, Epidermal Growth Factor ; Recurrence ; Retreatment ; Pemetrexed

A 5 year-old, intact female Yorkshire terrier was referred for dysuria and dyschezia. The radiographic and ultrasound examination showed a round shaped mass caudal to the urinary bladder that contained anechoic fluid within the thin walls. During surgery, the cyst was noted to be attached to the outer wall of the vagina, not connected to the vaginal lumen. Cystic fluid was removed and the cystic wall was resected. Then the remaining cystic wall was omentalized to prevent a recurrence. Histological examination confirmed that the cyst was of Wolffian duct origin. In this case, a large Gartner duct cyst causing urological problems was diagnosed and removed by surgical resection.
Animals ; Constipation/etiology/veterinary ; Cysts/surgery/ultrasonography/*veterinary ; Dog Diseases/*pathology/surgery/ultrasonography ; Dogs ; Dysuria/etiology/veterinary ; Female ; Treatment Outcome ; Vaginal Diseases/complications/pathology/surgery/*veterinary ; Wolffian Ducts/*pathology/surgery

The majority of flexible bronchoscopies are performed under topical anesthesia with lidocaine being the most commonly used agent. Anaphylaxis rarely occurs after local administration of lidocaine, but can be a fatal complication. We experienced a case of unexpected anaphylaxis. A 66-year-old woman was scheduled for flexible bronchoscopy to evaluate a tracheal mass and stenosis. The oral and nasal mucosa were pretreated with lidocaine. About 2~3 minutes later, the patient developed hypotension and we treated for anaphylaxis in the emergency room. Then, we decided to perform rigid bronchoscopy in this patient, under conditions of general anesthesia. A rigid bronchoscopy was performed in this patient, safely and successfully. The tracheal mass was determined to be squamous cell carcinoma.
Aged ; Anaphylaxis ; Anesthesia ; Anesthesia, General ; Bronchoscopy ; Carcinoma, Squamous Cell ; Constriction, Pathologic ; Emergencies ; Female ; Humans ; Hypotension ; Lidocaine ; Nasal Mucosa

The tools for asthma control assessment recommended by the current guideline are cognitive function- and effort-dependent, which is substantially impaired in the elderly. The aim of this study is to investigate objective assessment tools of asthma control status and previous asthma exacerbation (AE) in elderly subjects. Asthmatics aged >60 years who were treated with step 2 or 3 by the Global Initiative for Asthma (GINA) guideline were enrolled. During the 12-week study period, the subjects used either 400 µg of budesonide plus 10 mg of montelukast or 800 µg of inhaled budesonide. The occurrence of AE during the 4-week run-in and 12-week treatment period was monitored. After 12-week of treatment, sputum eosinophil count, peripheral eosinophil count, the plasma leukotriene E₄ (LTE₄), and prostaglandin F₂α (PGF₂α) metabolite levels were measured using the UHPLC/Q-ToF MS system. The study subjects were divided into group 1 (asthmatics who experienced AE during the study period) and group 2 (those who did not). A total of 101 patients aged 60-85 years were enrolled. Twenty-three patients (22.8%) had experienced AE. The plasma LTE₄ level, LTE₄/PGF₂α ratio, and peripheral eosinophil count were significantly higher in group 1 than in group 2 (P=0.023, P=0.010, P=0.033, respectively). The plasma LTE₄/PGF₂α ratio and peripheral eosinophil count at week 12 were significantly associated with previous AE (odds ratio [OR]=1.748, P=0.013; OR=1.256, P=0.027). Receiver operating characteristic (ROC) curves to discriminate the subjects with previous AE, including these 2 parameters, showed that the area under the curve was 0.700 (P=0.004), with 73.9% sensitivity and 47.9% specificity. In conclusion, a combination of plasma LTE₄/PGF₂α ratio and peripheral eosinophil count can be an objective assessment tool which is significantly associated with asthma control status in elderly asthmatics.
Aged* ; Asthma* ; Budesonide ; Eosinophils* ; Humans ; Plasma* ; ROC Curve ; Sensitivity and Specificity ; Sputum