Background : Recently the incidence of tuberculosis is increasing in many countries and control of the disease is further threatened by the emergence of multi-drug resistant tuberculosis. So rapid detection of drug resistance is very important. Pyrazinamide (PZA) is a first-line chemotherapeutic agent for tuberculosis. Now in Korea, we perform PZase activity test instead of actual pyrazinamide susceptibility test for the detection of PZA resistant M. tuberculosis. Recently the pncA gene, encoding the PZase of M. tuberculosis, was completely sequenced. And it was reported that the mutation of pncA gene would be associated with PZA resistance of M. tuberculosis. Therefore we performed this study to evaluate the possibility for the rapid detection of PZA resistant M. tuberculosis using PCR-SSCP of pncA gene. Method : 44 cultured clinical isolates of M. tuberculosis, BCG Tokyo strain. BCG French strain, and one M. bovis isolate were studied. We used H37Rv as the reference strain, The PZase activity test was done at the reference laboratory of Korean Tuberculosis Institute. DNA was extracted by bead-beater method and 561 bp fragment including pncA gene was amplified by PCR. The PCR product were digested by BstB I enzyme. SSCP was done using MDE gel. Of the 44 strains of M. tuberculosis, 22 strains were PZase-positive and other 22 strains were PZase negative. Results : Of the 22 PZase positive strains, 18 strains(82%) showed the same mobility compared with that of H37Rv and 4(18%) showed different mobility. Of the 22 PZase-negative strains, 19(86%) strains showed the same mobility pattern compared with that of H37Rv and 3(14%) showed different mobility. Naturally PZA-resistant BeG-French strain, BCG-Tokyo strain, and one M. bovis isolate showed the same band pattern each other, but their mobility were different from that of H37Rv. The results of PZase activity test and PCR-SSCP of pncA of M. tuberculosis were statistically significantly correlated each other (p<0.01). Conclusion : The PCR-SSCP after BstB I restriction of pncA gene of M. tuberculosis may be a useful method for the rapid detection of PZA-resistant M. tuberculosis.
Park, Sam-Seok;Kwak, Kyung-Rok;Hwang, Ji-Yun;Yun, Sang-Myeong;Ryue, Chi-Chan;Chang, Chul-Hun;Lee, Min-Gi;Park, Sun-Gue
Tuberculosis and Respiratory Diseases
/
v.47
no.6
/
pp.747-756
/
1999
Background: Acid-fast stain and cultures for diagnosis of pulmonary tuberculosis are primary and essential method, but have their limitation : low sensitivity and time consuming. The objective of this study is comparison of amplified Mycobacterium tuberculosis direct test(MTD) by the conventional AFB smears and cultures in the detection of Mycobacterium tuberculosis in respiratory specimens. Methods: During the period between November, 1997 and May, 1998 a total of 267 respiratory specimens (sputum 173, bronchial washing 94) from 187 patients suspected pulmonary tuberculosis were subjected to AFB smears, cultures and MID test. MID is based on nucleic acid amplification. We compared the MID with 3% Ogawa culture method. In positive AFB smear and negative MID specimen, positive culture identification between nontuberculous mycobacterium and M.tuberculosis was assesed by using Accuprobe M.tuberculosis complex probe. In negative AFB smear and negative AFB culture, MTD results are assessed by clinical follow-up. Results : 1) Compared with culture in sputum and bronchial fluid specimens, sensitivity and specificity of MTD in positive AFB smear is 79.7% and 20.0%, sensitivity and specificity of MTD in negative AFB smear specimens is 75.0% and 79.7%. 2) Discrepant analysis is assessed by clinical follow-up and other specimen results beyond study. Culture negative but MTD positive specimens were proved to be true positive and gave MTD sensitivity 79.2%, specificity of 84.4%, positive predictive value 80.5% and negative predictive value 83.2%. 3) 14 out of 31 specimens in negative AFB smear, negative AFB culture and positive MTD showed pulmonary tuberculosis diagnosed on clinical follow-up and sensitivity is 45.2%. 4) 2 out of 13 specimens in positive AFB smear, positive AFB culture and negative MID diagnosed as non tuberculous mycobacterium by Accuprobe culture. Conclusion: This study suggested that MID in respiratory specimens is simple and rapid diagnostic method, but considered adjuvant method rather than replace the conventional AFB smear and culture.
Lee, Jung Yeon;Choi, Hee Jin;Lee, Hyeyoung;Joung, Eun Young;Huh, Jin Won;Oh, Yeon-Mok;Lee, Sang Do;Kim, Woo Sung;Kim, Dong Soon;Kim, Won Dong;Shim, Tae Sun
Tuberculosis and Respiratory Diseases
/
v.58
no.4
/
pp.385-391
/
2005
Background : The incidence of nontuberculous mycobacterium (NTM) infections in Korea is increasing. This retrospective study was performed to examine the recovery rate of NTM from respiratory specimens as well as the isolated NTM colony characteristics, and to assess the clinical significance of a NTM isolation. Methods : The results of the respiratory specimens requested for an acid-fast bacilli (AFB) examination during 2002 at Asan Medical Center, along with the patients clinical characteristics were analyzed. Results : A total 26,820 respiratory specimens were requested for the acid-fast bacilli (AFB) smear and culture during the study period. The proportion of M. tuberculosis and NTM isolation was 5.7% and 2.2%, respectively. Among the AFB smear and culture positive specimens, 12.2% were found to be NTM. The scotochromogen showing a low colony count < 20, which appeared to be contaminants, were isolated in 31.8% of the 584 NTM isolates. Excluding the low-colony scotochromogens, the M. avium-intracellulare complex was the most common NTM isolates (42.1%), and was also the most common causative organism for NTM pulmonary diseases. 8.4% (23/275) and 17.8% (49/275) of patients with NTM isolates met the American and British Thoracic Society diagnostic criteria for NTM pulmonary disease, respectively. Conclusion : In case of a positive AFB-smear or culture result, the possibility of NTM being a causative organism should always be considered, even in Korea, which has an intermediate incidence of tuberculosis.
Koo, Hee Soo;Ku, Pyeong Tae;Lee, Mi Ok;Baik, Hyung Suk
Journal of Life Science
/
v.26
no.8
/
pp.911-920
/
2016
Norovirus is the most common causative agent of acute gastroenteritis. This study was carried out to investigate molecular epidemiology of norovirus infections from outbreaks in Busan from 2012 to 2015. Total of 581 stool specimens were collected from diarrhea patients in outbreaks in Busan, 71 samples were resulted in positive to norovirus. The data were analyzed according to seasonality, patient, age and gender. Noroviruses were detected most frequently during the winter season from November (25.4%) to February (28.2%). The age group from teens was the most susceptible to norovirus infections. To obtain the molecular genetic information of norovirus, we performed sequencing analyses of the strains detected. Norovirus genotypes have been reported to show high genetic diversity. Four kinds of GI genotypes (GI-1, GI-2, GI-3, GI-5) and five kinds of GII genotypes (GII-1, GII-4, GII-5, GII-6, GII-17) were indentified in outbreaks in Busan. Other previous studies have shown that GII-4 is the most predominant circulating in Korea and worldwide. The most prevalent norovirus genotypes of each year were GII-6 in 2012, GII-6 in 2013, GII-4 in 2014 and GII-6 in 2015. Except for 2014, GII-6 genotype was the most prevalent and predominant in Busan. We described the epidemiological analysis of the noroviruses in outbreaks in Busan. The result of this study will contribute to update the epidemiological data and improve hygiene and public health via sustainable surveillance.
Chun Mison;Kang Seunghee;Jin Yoon-Mi;Oh Young-Taek;Kil Hoon-Jong;Oh Tae-Young;Ahn Byoung-Ok
Radiation Oncology Journal
/
v.19
no.3
/
pp.265-274
/
2001
Purpose : Proctitis is one of acute complications encountered when radiotherapy was appled to the pelvis. Radiation-induced proctitis represents similar microscopic findings that are observed in inflammatory bowel disease (IBD). Nitric oxide (NO) plays an important role in the inflammatory process and many data suggest a close relationship between NO production and gastrointestinal inflammation. This study was aimed to establish the optimal radiation dose for radiation-induced proctitis in rat and to find a relationship between radiation proctitis and NO production. Materials and methods : Female Wistar rats, weighing from 150 to 220 g, received various doses(10-30 Gy) of radiation to the rectum. On the 5th and 10th day after irradiation, rectal specimens were evaluated grossly and microscopically. In addition, the degree of NO production by irradiation dose was evaluated by study with NOS expression and nitrite production in the irradiated rectal tissue. To evaluate relationship between radiation proctitis and NO, we administered aminoguanidine, iNOS inhibitor and L-arginine, substrate of NOS to rats from 2 days before to 7 days after the irradiation. Results : There were obvious gross and hostological changes after 17.5 Gy or higher radiation dose but not with 15 Gy or less radiation dose. Twenty Gy or higher dose of radiation caused Grade 4 damage in most of rectal specimens which were more likely to be related to the late complications such as fibrosis, rectal bleeding and rectal obstruction. A single fraction of 17.5 Gy to the rat rectum is considered to be an optimal dose to produce commonly experienced proctitis in the clinic. The result demonstrated that severity of microscopic damage of rectal mucosa from irradiation significantly correlated with iNOS over-expression. However, administration of iNOS inhibitor or substrate of iNOS did not influence the degree of rectal damage. Conclusion : A single fraction of 17.5 Gy irradiation to the rat rectum considered to be an optimal dose for radiation induced proctitis model. These results indicated that an excess production of NO contributes to pathogenesis of radiation-induced proctitis in part but was not the direct cause of rectal damage.
Purpose: It is known that lactoferrin serves as a source of iron for H. pylori in gastric mucosa. This study was undertaken to investigate the relationship between lactoferrin and H. pylori infection coexistent with iron-deficiency anemia by determining the lactoferrin levels in gastric biopsy specimens, and by locating the major sites of lactoferrin expression, according to the presence or absence of iron-deficiency anemia. Methods: Fifty-five adolescents that underwent gastroduodenoscopy were divided into three groups: NL (n=19) for normal controls, HP (n=15) for patients with H. pylori, and IDA (n=21) for patients with H. pylori gastritis and coexisting iron-deficiency anemia. Histopathologic features were graded from to marked on the basis of the Updated Sydney System. The gastric mucosal levels of lactoferrin were measured by immunoassay. Immunohistochemical technique was used to allow identification of the location and quantification of the lactoferrin expression. Results: Lactoferrin levels in the antrum increased significantly, in proportion to, H. pylori density, polymorphonuclear cell infiltration, and chronic inflammation in the histologic specimens. Patients in the HP and IDA groups showed significantly increased mucosal levels of lactoferrin compared with that observed in the normal group (p=0.0001). The lactoferrin level in IDA group tended to be higher than that in the HP group (p=0.2614). The major sites of lactoferrin expression by immunohistochemistry were in glands and neutrophils within epithelium. Lactoferrin was stained weakly in NL, and strongly in HP and IDA. Conclusion: The lactoferrin sequestration in the gastric mucosa of IDA was remarkable, and this finding seems to give a clue that leads to the clarification of the mechanism by which H. pylori infection contributes to iron-deficiency anemia.
Purpose: To prevent residual physical disability and chronic infection, prompt diagnosis and adequate treatment are important in the skeletal infections in children. Although radioisotope scanning is knwon as the method of choice for early diagnosis of bone infection, we conducted a study on twenty nine children who had skeletal infections to reevaluate the most appropriate way in diagnosis and management. Methods: A retrospective study was conducted on twenty nine children, who were admitted to the departments of Pediatrics and Orthopedic Surgery and who had acute osteomyelitis or septic arthritis, through review of medical records, radiologic & radioisotope study results. Their diagnoses were confirmed by bacteriologic cultures on the aspirated specimens from suspected bony lesions. Results: 1) Among twenty nine patients, there were 6 infants including 5 newborn infants, and 23 children were aged between 1 and 15 years. Male to female ratio was 1.4 to 1. 2) Point tenderness was noted in all cases, and the common physical signs were swelling, limitation of motion, fever and local heat in the order of frequency. 3) Fifty two percents of the patients were diagnosed within a week after onset of symptoms and all cases were within 15 days. 4) Leukocytosis was noted in only 58.6% of cases but erythrocyte sedimentation rate was increased in all cases except only one case. Staphylococcus aureus was revealed as the most common etiologic agent. 5) Radioisotope scans showed hot uptake in five of six cases(83.3%) who had no abnormal finding on plain skeletal radiolograms. Conclusions: Although radioisotope scan and MRI are helpful in early diagnosis before radiologic finding was detected on plain X-ray film, the antimicrobial therapy can be started after bacteriologic study of the aspirated specimens from the suspected skeletal lesions if skeletal infection is highly suspected clinically.
Kim, Nam Hee;Lee, Jina;Lee, Jung Won;Lee, Soo Young;Choi, Eun Hwa;Kim, Kyung Hyo;Kim, Eui Chong;Kang, Jin Han;Lee, Hoan Jong
Pediatric Infection and Vaccine
/
v.16
no.1
/
pp.31-39
/
2009
Purpose : Neisseria meningitides is one of the most common causative pathogens of bacteremia and meningitis. Recently protein-conjugated vaccines have been developed and included in the routine vaccination schedule in a few countries. In Korea, carriage rates of N. meningitides among healthy adults have been reported. However, systematic data for childhood carriage rates are not available. This study was performed to evaluate the carriage rates of N. meningitides and the serotype distribution among healthy children attending day care centers. Methods : During the period of January through May 2005, nasopharyngeal swabs and culture were obtained from 904 children attending 13 different day care centers located in Seoul and Gyeonggi Province. The Vitek NHI card was used to identify N. meningitides and the crgA gene was detected via polymerase chain reaction (PCR). Serotype determination was performed by agglutination test using N. meningitides antisera to serotypes A, B, C, D, 29E, W135, X, Y, and Z. PCR for detection of the org2 and saiD gene confirmed serotypes A, B, C, W135, and Y. Results : The mean age among 904 children was 4.5 years; 6.5% (59/904) were children <2 years old, 53.8% (486/904) were 2-5 years old, and 39.7% (359/904) were >5 years old; 52.0% (468/904) were male. N. meningitides was isolated from only 7 children attending 5 different day care centers and the overall carriage rate of N. meningitides was 0.8%. The detected serotypes of N. meningitides were serotype A (n=2), C (n=2), and Y (n=3). Conclusion : The carriage rate of N. meningitides among healthy children attending day care centers was very low in Korea and the detected serotypes were A, C, and Y.
Kim, Seung Yun;Lee, Ko Eun;Kang, Su Young;Choi, Eun Hwa;Lee, Hoan Jong
Pediatric Infection and Vaccine
/
v.22
no.3
/
pp.172-177
/
2015
Purpose: This study aimed to analyze the epidemic period of RSV infection and evaluate the appropriate time of palivizumab immunoprophylaxis. Methods: From January 1991 to July 2012, nasopharyngeal (NP) aspirates were obtained from patients who visited Seoul National University Children's Hospital for respiratory symptoms. NP samples were used to detect respiratory viruses. Among them, we analyzed the positive number and detection rate of RSV infection in two-week interval. The beginning of RSV season was defined when RSV positive number was more than 4 and RSV detection rate was over 10%. From January 2007 to March 2014, we analyzed the starting time of palivizumab immunoprophylaxis for the infants at high risk. Results: The RSV detection rate was 2,013/21,698 (9.69%) over 22 years. The median RSV season was from $2^{nd}-3^{rd}$ week of October to $1^{st}-2^{nd}$ week of February. The earliest starting week was the 3rd week of July in year 2001, and the latest end week was the 3rd week of May in year 1990. Palivizumab immunoprophylaxis was initiated most frequently at the 3rd week of October (18.7%). However, the percentage of starting palivizumab on the 1st week of September has increased from 3.8% in the year 2007 to 14.1% in 2013. Conclusions: The year to year variability of RSV season exists. The starting time of palivizumab immunoprophylaxis should be adjusted based on the season of RSV epidemic.
Purpose: Although ${\alpha}$-fetoprotein is one of the most commonly used tumor markers in Korea, most of the radioimmunoassay kits for ${\alpha}$-fetoprotein have been imported from foreign countries. The purpose of this study was to evaluate the performance of a recently developed domestic immunoradiometric kit for ${\alpha}$-fetoprotein (Riakey AFP IRMA $CT^R$, Sin-Jin Medics, Seoul, Korea). Materials and Methods: We evaluated intra- and inter-assay precision, recovery rate, parallelism, and sensitivity of serum ${\alpha}$-fetoprotein measurement using Riakey AFP IRMA $CT^R$ kit. The values of ${\alpha}$-fetoprotein measured by Riakey AFP IRMA $CT^R$ kit were compared with those measured by two foreign commercial kits (${\alpha}$-fetoproteina of Radim and ${\alpha}$-feto.riabead of Abbott). Results: Intra-assay coefficients of variation on three different levels were 5.3% for 18.9 ng/ml, 3.4% for 133 ng/ml and 1.6% for 330 ng/ml. Inter-assay coefficients of variation were 9.7% for 20.9 ng/ml, 3.2% for 137 ng/ml and 4.1% for 330 ng/ml respectively. Recovery rate tests on all three different levels showed within $100{\pm}10%$. Parallelism was also good and the sensitivity was 0.63 ng/ml. There was strong correlation between the measurement of ${\alpha}$-fetoprotein by Riakey AFP IRMA $CT^R$ and that by two foreign commercial kits(r=0.98). Conclusion: The first Korean domestic immunoradiometric kit for ${\alpha}$-fetoprotein, Riakey AFP IRMA $CT^R$, performed well for clinical use.
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