BACKGROUND: HLA-B*15 alleles encode molecules belonging to several serologic subtypes, B62, B63, B71, B72, B75, B76, and B77. Using the conventional serologic typing method, assignment of B15 subtypes has often been prone to error specifically in samples exhibiting either an ambiguous or a B15 homozygous reaction pattern. The goal of this study was to establish a supplementary DNA typing method for accurate assignment of B15 subtypes in 'problematic B15 positive samples'. METHODS: B*15 specific gene amplification was performed using a pair of PCR primers that specifically annealed to B*15 and B*46 alleles. Nested PCR was applied to the amplified DNA using 14 sequence specific PCR primer sets. DNA sequencing was used to clarify the assigning of samples exhibiting discrepancies between the results obtained by B*15-specific nested PCR-SSP typing and serology. RESULTS: The B*15-specific nested PCR-SSP typing could clearly discriminate the 9 B*15 alleles expressed in the Korean population. In application of the system to 30 B15 positive serologically typed samples, 4 exhibited discrepancies between serology to PCR-SSP results. DNA sequencing results obtained from the samples were concordant with those from B*15-specific nested PCR-SSP typing. CONCLUSIONS: The established B*15-specific nested PCR-SSP method is superior to serology in accuracy and resolution. Therefore, the method will be useful as a supplementary DNA typing method to clarify HLA-B assignments of 'problematic B15 positive samples' in Koreans.
Alleles ; DNA Fingerprinting* ; DNA* ; Gene Amplification ; HLA-B Antigens ; Polymerase Chain Reaction ; Sequence Analysis, DNA

BACKGROUND: Early diagnosis of acute myocardial infarction (AMI) is essential for the patient's clinical course. Serum cardiac markers such as creatine kinase MB (CK-MB), myoglobin, and cardiac troponin I (cTnI) supply physicians with powerful methods of diagnosing AMI. We evaluated the usefulness of ACS:CentaurTM for the tests of cardiac markers. METHODS: We used three levels of control materials and 138 patient samples (AMI 49, cardiac diseases other than AMI 41, noncardiac diseases 48). We analyzed precision and linearity of CK-MB, myoglobin and cTnI using control materials. The cutoffs for the descrimination of AMI were determined using receiver operating characteristic (ROC) curve, and sensitivity and specificity of each marker were analyzed. RESULTS: Coefficients of variations (CVs) of within day precision and between day precision were below 4% except cTnI. Linearity analyses showed good results in all the three markers. The cutoffs were 3.7 ng/mL for CK-MB, 121.0 ng/mL for myoglobin and 0.4 ng/mL for cTnI. In the diagnosis of AMI, the sensitivities and specificities were 93.9% and 97.8% for CK-MB, 73.5% and 91.0% for myoglobin and 100.0% and 95.5% for cTnI, respectively. CONCLUSIONS: The tests of cardiac markers using ACS:CentaurTM showed satisfactory precision and linearity, and they are specific and sensitive in the diagnosis of AMI. So, ACS:CentaurTM is useful for the measurement of cardiac markers in the diagnosis of AMI.
Creatine Kinase ; Diagnosis ; Early Diagnosis ; Heart Diseases ; Humans ; Myocardial Infarction ; Myoglobin ; ROC Curve ; Sensitivity and Specificity ; Troponin I

Providencia stuartii is occasionally isolated from patients with chronic otitis media, however it has not been reported as a cause of acute otitis media so far. We recently isolated P. stuartii from the pus specimen of right middle ear of a 2-year-old patient with acute otitis media. The patient was admitted because of right-sided otorrhea for 2 weeks. The symptom was not relieved by first-line empirical antibiotics (amoxicillin-clavulanic acid and netilmicin), but it was subsided dramatically with the changed antibiotics (ceftriaxone and amikacin); the isolate was susceptible to these two antibiotics in in vitro susceptibility testing. To our knowledge, this is the first case report of acute otitis media caused by P. stuartii.
Anti-Bacterial Agents ; Child, Preschool ; Ear, Middle ; Humans ; Otitis Media* ; Otitis* ; Providencia* ; Suppuration