Nghiem, Minh Ngoc;Nguyen, Bac Van;Nguyen, Son Thai;Vo, Thuy Thi Bich;Nong, Hai Van
Journal of Microbiology and Biotechnology
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v.25
no.5
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pp.745-752
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2015
Tuberculosis (TB) is the most common mycobacterial infection in developing countries, requiring a rapid, accurate, and well-differentiated detection/diagnosis. For the rapid detection and discrimination of Mycobacterium tuberculosis complex (MTC) from non-tuberculous mycobacteria (NTM), a novel, simple, and primer-combined single-step multiplex PCR using three primer pairs (6110F-6110R, 1081F-1081R, and 23SF-23SR; annealing on each of IS6110, IS1081, and 23S rDNA targets), hereafter referred to as a triplex PCR, has been developed and evaluated. The expected product for IS6110 is 416 bp, for IS1081 is 300 bp, and for 23S rDNA is 206 bp by single PCR, which was used to verify the specificity of primers and the identity of MTC using DNA extracted from the M. tuberculosis H37Rv reference strain (ATCC, USA) and other mycobacteria other than tuberculosis (MOTT) templates. The triplex PCR assay showed 100% specificity and 96% sensitivity; the limit of detection for mycobacteria was ~100 fg; and it failed to amplify any target from DNA of MOTT (50 samples tested). Of 307 blinded clinical samples, overall 205 positive M. tuberculosis samples were detected by single PCR, 142 by conventional culture, and 90 by AFB smear methods. Remarkably, the triplex PCR could subsequently detect 55 positive M. tuberculosis from 165 culture-negative and 115 from 217 AFB smear-negative samples. The triplex PCR, targeting three regions in the M. tuberculosis genome, has proved to be an efficient tool for increasing positive detection/discrimination of this bacterium from clinical samples.
Im, Bong Chil;Kim, Young;Kim, Kyoung Sim;Kim, Yong Wook;Kim, Eun Young;You, Eun Jung;You, Ju Hee;Cho, Hyoung Min
Pediatric Infection and Vaccine
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v.17
no.2
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pp.148-155
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2010
Purpose : The purpose of this study was to investigate clinical features and culture-positive rates according to the involved lung in adolescent pulmonary tuberculosis (TB). Methods : We retrospectively reviewed the medical records of adolescents who ranged in age from 10 to 20 years and who had been hospitalized with a diagnosis of TB at Kwangju Christian Hospital from 2000 to 2008. Results : Sixty-six patients were identified with pulmonary TB: median age 16.82 years; 48.5% males. Among them, 90.9% of patients were between 15 and 20 years of age. Most patients presented with multiple symptoms, and the most common included cough (74.2%), sputum (60.6%), fever (39.5%), and night sweating (18.2%). Sputum samples were smear-positive in 28 (42.4%), culture-positive in 40 (60.6%), and PCR-positive in 46 (69.7%). The most common radiological patterns included cavitation in 18 (27.3%), pleural effusion in 18 (27.3%), lymphadenopathy in 10 (15.2%), and tuberculoma in 5 (7.6%). The prevalence of smear, culture, and PCR positive rates increased as the number of involved lobes increased (P<0.05, P<0.01, P<0.05). The median treatment duration was 7 months. Twelve patients (18.2%) had lower lung field TB (Group A) and forty-four patients (66.7%) had other areas involving TB, except for Group A (Group B), and ten patients (15.1%) had only TB pleurisy (Group C). The difference of clinical characteristics and culture rates between group A and group B was not significant. Conclusion : Pulmonary TB toward late adolescence is increasing. We need to pay more attention to lower lung field TB, which is difficult to detect with specific radiographic findings.
Kim, Eun Kyung;Shim, Tae Sun;Lee, Jung Yeon;Oh, Yeon-Mok;Lim, Chae-Man;Lee, Sang Do;Koh, Younsuck;Kim, Dong Soon;Kim, Won Dong;Kim, Woo Sung
Tuberculosis and Respiratory Diseases
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v.57
no.3
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pp.226-233
/
2004
Background : Interferon-gamma (IFN-${\gamma}$) is a critical cytokine in the defense against a Mycobacterium tuberculosis infection. Even though IFN-${\gamma}$ has occasionally been used in the treatment of refractory multidrug-resistant tuberculosis (MDR-TB) with some promising results, there is still some controversy regarding the therapeutic efficacy of IFN-${\gamma}$. This study was performed to examine the effect of subcutaneous IFN-${\gamma}$ in the treatment of MDR-TB patients. Methods : Six patients with refractory MDR-TB were enrolled in this study. Two million IU of IFN-${\gamma}$ was administered subcutaneously three times a week with the concomitant administration of antituberculous drugs for at least for 28 weeks. During the IFN-${\gamma}$ therapy, the sputum smear and culture, radiological and clinical evaluations were performed every 4 weeks throughout the study period. Results : The mean age of the 6 patients was 37 years (ranges, 15-61 years). The drug susceptibility test to standard antituberculous drugs revealed resistance to an average of 6.8 (${\pm}1.2$) agents including isoniazid and rifampicin. An average of 10.8 (${\pm}1.3$) antituberculous drugs were prescribed before IFN-${\gamma}$ therapy. The culture became negative in 2 patients (33%) after initiating IFN-${\gamma}$ therapy; one at 8 weeks, and the other at 24 weeks. Finally, after stopping the IFN-${\gamma}$ therapy after 28 weeks, the culture became positive again in the two patients who were culture-negative. The other 4 patients who failed in the culture conversion are still on antituberculous treatment except for one who died of tuberculosis. Conclusion : Even though 28 weeks of subcutaneous IFN-${\gamma}$ therapy in combination with antituberculous drugs was successful in inducing the culture-negative conversion in some patients with refractory MDR-TB, the culture became positive again after stopping the IFN-${\gamma}$ therapy. This suggests that subcutaneous IFN-${\gamma}$ therapy may have suppressive effect on tuberculosis only during the IFN-${\gamma}$ therapy period in some patients. Further studies will be needed to determine the optimum dose, the administration route, the duration of therapy, and the predicting factors of the response to adjuvant IFN-${\gamma}$ therapy.
Background: Several potential risk factors have been identified for diffuse large B-cell lymphoma (DLBCL); however, epidemiological studies investigating the association between these risk factors and DLBCL have yielded inconsistent results. Objectives: To investigate potential medical, lifestyle, and environmental risk factors of DLBCL in Shanghai, China through a hospital-based case-control study. Method: One-hundred-and-forty-seven newly diagnosed DLBCL patients and 294 sex- and age-matched controls were recruited from 11 hospitals in Shanghai between 2003 and 2007. A standardized structured questionnaire was used to obtain patient data on demographics, medical history, family history, lifestyle, and environmental exposures. Conditional logistic regression models were used to estimate odds ratios (ORs), with 95% confidence intervals (CIs), for risk associated with each data category. Results: History of tuberculosis (TB) infection and "living on a farm" were positively associated with DLBCL (TB: OR=3.05, 95% CI: 1.19-7.80; farm: OR=1.82, 95% CI: 1.21-2.73). In contrast, taking traditional Chinese medicine was negatively associated with DLBCL (OR=0.36, 95% CI: 0.14-0.89). No significant correlation with DLBCL risk was found for any of the other potential risk factors (p>0.05), including but not limited to hair dyes, alcohol drinking, smoking, and home/workplace renovation within one year. Conclusions: Consistent with results from previous studies in other DLBCL case populations, traditional Chinese medicine appeared to have a direct or indirect protective effect against DLBCL. However, this study also identified a possible predisposition for DLBCL in TB sufferers and farmers.
1. The sixty raised shepherd and sixty-five inhoused pet dogs in the regions of Daejon and Cheongju were subjected to investigate the TB infection by means of BCG and X-ray diagnosis. The 5 out of 65 inhoused pet and 7 out of 60 shepherd dogs were observed to be infected with TB, respectively. However, none of Mycobacterium species were detected from lung tissues of 4-slaughtered dogs showing BCG positive reaction. 2. The rats were first inoculated with 0.1ml BCG, and then 0.1ml M bovis suspended solution($1{\times}10^5$ organisms/0.2ml) 3weeks later. After 5 months, the animals were killed. The pathohistological results from both groups, TB inoculated and BCG treated groups, were observed on the surface of lung. Furthermore, the severe pathological lesion in the Iung was observed in M bovis inoculated group compared to BCG treated group. 3. The slight macrophage invasion and granuloma formation in the lung from BCG treated group were observe individually. However, it was confirmed that the lung from M bovis treated group was invaded by the macrophages and neutrophils combined with the granuloma formation. 4. When the numbers of the total cells taken from broncho-alvealar fluid in each of mouse from both groups were differentially counted, the number of total cell, neutrophils, and lymphocytes from M bovis treated group were significantly increase compared with those of BCG treated group. 5. Although there were nearly no response of the alveolar macrophages to CSF in serum obtained from control group, those from M boris treated group were significantly proliferated.
Hong, Sung Wan;Hong, Chan Eui;Lee, Dong Jean;Roh, Byung Hak
Pediatric Infection and Vaccine
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v.15
no.2
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pp.174-179
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2008
Purpose : The purpose of this study was to investigate the clinical features of adolescent TB. Methods : We retrospectively reviewed the medical records of the adolescents aged 12 to 18 years and who are hospitalized with the diagnosis of TB at Dong Kang General Hospital between January 2003 and December 2007. Results : Of the 29 patients who were identified, 65.5% were male. The median age at diagnosis was 16.9 years (range: 12.1-18.9 years). A tuberculin skin test (TST) was performed for 6 patients (20.7%). Among them, 5 (5 of 8[62.5%]) in the 12-15 years age group (Group A) and 1 (1 of 21[4.8%]) in the 16-18 years of age group (Group B) were TST positive, which showed that the TST performing rate decreased with age (P<0.01). Twenty-five patients (86.2%) were detected by evaluating the clinical symptoms that suggested TB and 4 patients (13.8%) were detected by screening, but no cases were detected by contact tracing. Twenty patients (69.0%) had isolated pulmonary disease. Seven patients (24.1%) had pulmonary and extrapulmonary disease and 2 patients (6.9%) had exclusively extrapulmonary disease. Most patients presented with multiple symptoms, and the most common were cough (79.3%), fever (75.9%), sputum (65.8%) and chest pain (34.5%). M. tuberculosis was isolated from the sputum of 20 patients (69.0%). The culture-positive rate increased with age (range: 62.5% in Group A to 71.4% in Group B), but the difference between the groups was not significant. Cavitary lesions were found 13 patients (44.8%). The cavitary lesion rate increased with age (range: 25.0% in Group A to 52.4% in Group B), but the difference between the groups was not significant. Four patients (13.8%) were lost during follow-up. One patient (6.9%) relapsed. Conclusion : Clinicians need to be aware of the unique features of adolescent TB, and they also need awareness of the importance of TST and contact investigation for an adult suspected of having TB for making the diagnosis of TB.
Chun, Jin-Kyong;Kim, Chang Ki;Kim, Hyun Sook;Jung, Ghee Young;Linton, John A.;Kim, Ki Hwan;Lee, Taek Jin;Jeon, Ji Hyun;Kim, Dong Soo
Clinical and Experimental Pediatrics
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v.51
no.9
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pp.971-976
/
2008
Purpose : Surveillance for detecting and managing latent tuberculosis infection (LTBI) is a key component of tuberculosis control. The classic surveillance tool, the tuberculin skin test (TST), may have some limitations when used in the Bacillus Calmette-$Gu{\acute{e}}rin$ (BCG)-vaccinated population. The object was to perform a blood test $QuantiFERON^{(R)}$-TB Gold In Tube (QFT-G IT) based on the detection of interferon-$\gamma$ ($IFN-{\gamma}$) released by T cells in response to Mycobacterium tuberculosis-specific antigens, and to compare the efficacy of this new diagnostic tool for LTBI with that of TST. Methods : For six months, between October 1, 2006 and April 30, 2007, data were collected from 111 patients under 15 years of age at Severance Children's Hospital. TST and QFT-G IT tests were performed with children with or without contact histories of tuberculosis. In addition to these tests, we examined comparative data from 29 adults who had tuberculosis, to detect false negative rates in the QFT-G IT method. Results : Thirty-three children had household contact histories. In this group, 15% and 42% of cases were found to be positive using the QFT-G IT assay and TST, respectively. Agreement was low between these two tests (${\kappa}=0.39$). In the adult active tuberculosis group, the QFT-G IT false negative rate defined as a positive culture and a negative QFT-G IT result was 12.5%. Conclusion : In diagnosing LTBI in children, the usefulness of a whole-blood $IFN-{\gamma}$ assay employing TB-specific antigens will be revealed only by examining additional longitudinal clinical data; this study serves as a starting point in that process.
T cell exhaustion develops under conditions of antigen-persistence caused by infection with various chronic pathogens, such as human immunodeficiency virus (HIV) and myco-bacterium tuberculosis (TB), or by the development of cancer. T cell exhaustion is characterized by stepwise and progressive loss of T cell function, which is probably the main reason for the failed immunological control of chronic pathogens and cancers. Recent observations have detailed some of the intrinsic and extrinsic factors that influence the severity of T cell exhaustion. Duration and magnitude of antigenic activation of T cells might be associated with up-regulation of inhibitory receptors, which is a major intrinsic factor of T cell exhaustion. Extrinsic factors might include the production of suppressive cytokines, T cell priming by either non-professional antigenpresenting cells (APCs) or tolerogenic dendritic cells (DCs), and alteration of regulatory T (Treg) cells. Further investigation of the cellular and molecular processes behind the development of T cell exhaustion can reveal therapeutic targets and strategies for the treatment of chronic infections and cancers. Here, we report the properties and the mechanisms of T cell exhaustion in a chronic environment.
Jung, Jae Woo;Lee, Young Woo;Choi, Jae Cheol;Yoo, Seung Min;Lee, Hwa Yeon;Lim, Seoung Young;Shin, Jong Wook;Kim, Jae Yoel;Park, In Whn;Kim, Mi Kyung;Choi, Byoung Whui
Tuberculosis and Respiratory Diseases
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v.60
no.5
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pp.523-531
/
2006
Background : Cervical tuberculous lymphadenopathy is a very common disease with a similar incidence to pulmonary tuberculosis. Dendritic cells play a role of initial antigen presentation of this illness. Nevertheless, the precise role of these antigen-presenting cells according to the clinical features in unclear. The aim of this study was to determine the clinical implication of dendritic cell infiltration in the cervical lymph nodes. Methods : A review of the clinical characteristics was carried out retrospectively based on the clinical records and radiography. Immunohistochemical staining was performed on the available histology specimens of 72 cases using the S-100b polyclonal antibody for dendritic cells. The number of dendritic cells with tuberculous granuloma were determined. A $X^2$ test, unpaired T test and multiple logistic regression analysis were performed. Results : Thirty percent of subjects had previous or concurrent pulmonary TB. Twenty one percent of cases showed a positive reaction on the AFB stain. Within a granuloma, the number of infiltrated dendritic cells was $113.0{\pm}7.0$. The incidence of fever and cough decreased with increasing infiltration of dendritic cells Multivariate regression analysis showed that the infiltration of dendritic cells could significantly contribute to fever. Conclusion : Overall, dendritic cells can control a Mycobacterium tuberculosis infection and modulate the immune response, as well as resolve the clinical manifestations of TB lymphadenopathy.
Koo, Ho Seok;Kim, Tae Kyun;Park, Sung Kil;Choi, Sang Bun;Kim, Ae Ran;Choi, Sang Bong;Jung, Hoon;Park, I-Nae;Hur, Jin-Won;Lee, Hyuk Pyo;Yum, Ho-Kee;Choi, Soo Jeon;Choi, Suk-Jin;Lee, Hyun-Kyung
Tuberculosis and Respiratory Diseases
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v.63
no.3
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pp.268-272
/
2007
A tuberculous pleural effusion may be a sequel to a primary infection or represent the reactivation of pulmonary tuberculosis. It is believed to result from a rupture of a subpleural caseous focus in the lung into the pleural space. It appears that delayed hypersensitivity plays a large role in the pathogenesis of a tuberculous pleural effusion. We encountered a 52 years old man with pleural effusion that developed several days after a CT guided percutaneous needle biopsy of a solitary pulmonary nodule. He was diagnosed with TB pleurisy. It is believed that his pleural effusion probably developed due to exposure of the parenchymal tuberculous focus into the pleural space during the percutaneous needle biopsy. This case might suggest one of the possible pathogeneses of tuberculous pleural effusion.
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