[STUDY OF DIRECT TB-LAMP USING NON-CENTRIFUGAL SPUTUMS ABOUT EFFICIENCY FOR RAPID DIAGNOSIS OF TUBERCULOSIS].

OBJECTIVE: To evaluate the efficiency of the direct tuberculosis-loop-mediated isothermal amplification (TB-LAMP) assay by using non-centrifuged sputum samples. STUDY PERIOD AND METHODS: The study was conducted between June 2013 and February 2014. We collected 111 sputum samples from patients who had been radiographically diagnosed with tuberculosis and had not received any treatments for longer than 5 days. In the direct TB-LAMP assay, a loop-mediated isothermal amplification kit and 60-µL sputum samples were used. A direct smear microscopy test was used as the smear test. Then, the same sputum samples were processed with a CCE pretreatment reagent, and 100 µL of the solution samples were cultured by using the mycobacterial growth indicator tube (MGIT) culture method. RESULTS: Forty-six of the 111 samples were positive in the smear microscopy tests. All the smear-positive samples were positive in both the MGIT and direct TB-LAMP assay (100%). The mean positive detection time with the direct TB-LAMP assay was 13 minutes 55 seconds. Of 56 smear-negative and MGIT positive samples, 44 (78.6%) were judged to be positive using the direct TB-LAMP assay, with a mean positive detection time of 15 minutes 59 seconds. DISCUSSION: The direct TB-LAMP assay using non-centrifuged sputum samples was demonstrated to have a high detection rate and thus may be considered useful for rapid and effective tuberculosis diagnosis.

[Comparative study of the efficacy of the COBAS TaqMan and LAMP assay for the rapid diagnosis of tuberculosis].

UNLABELLED: OBJECTIVE; The COBAS TaqMan real-time polymerase chain reaction (PCR) assay (TaqMan assay) is a well-accepted and widely distributed molecular-based diagnostic test for tuberculosis. In the present study, we evaluated the efficacy of the LAMP assay (loopamp MTBC detection kit) as an alternative molecular-based diagnostic kit for tuberculosis, through comparison with the TaqMan assay. STUDY PERIOD AND METHODS: This study was conducted over a period of approximately 2 months, between May and July 2012. We collected 48 samples (43 sputum, 2 gastric fluid, 2 pleural fluid, and 1 pus fluid samples) from patients who had been diagnosed with tuberculosis through the culture method, but had not received any treatment for more than one week. All samples were processed using the CC-E pre-treatment reagent (Japan BCG) prior to performing the TaqMan and LAMP assay. For the TaqMan assay, 100 microL of supernatant was used after centrifugation at 1,000 rpm for 1 minute, whereas 60 microL of the precipitate in the same sample was used for the LAMP assay. RESULTS: In total, 23 out of 48 samples were identified as positive for tuberculosis according to smear microscopy tests, among which 15, 4, and 4 samples had smear test scores or 1+, 2+, and 3+, respectively. All the samples that tested positive in the smear test, regardless of the score, also tested positive in both the TaqMan and TB-LAMP assays (100%). Of the 25 smear-negative samples, we noted that 16 tested positive by the TaqMan assay (64%), whereas 20 tested positive by the LAMP assay (80%). DISCUSSION: Compared with the TaqMan assay, the LAMP assay showed a higher positive rate among the smear-negative samples. We believe that this is because substances in the samples acted as co-precipitating agents, resulting in the presence of a larger number of bacteria in the precipitates than in the supernatants. Thus, the findings indicate that the application of the LAMP method to precipitates obtained following CC-E pre-treatments may lead to prompt diagnosis of tuberculosis, with a level of sensitivity comparable to that of culture tests.

Association between episodes of upper respiratory infection and exacerbations in adult patients with asthma.

BACKGROUND: Asthma has several phenotypical features, including recurrent exacerbations and recurrent episodes of upper respiratory infection (URI). PURPOSE: A retrospective study was performed to identify the characteristics of adult patients with recurrent exacerbations of asthma, especially in association with recurrent episodes of URI. METHODS: Information was collected using a self-administered questionnaire given to 7070 patients in autumn-winter 2006, 4859 patients in spring-summer 2007, and 4452 patients in autumn-winter 2007. The patients reported the degree of symptoms and the frequency of febrile episodes of URI and exacerbations. Severe exacerbations were defined as a self-report of asthma-related hospitalization, an emergency department visit, or a requirement for systemic corticosteroids. Recurrent febrile URI and exacerbations were defined as two or more episodes within the previous 6 months. A Poisson regression model was used to identify the factors that were predictors of a risk for exacerbations. RESULTS: Of the 6266 patients who completed the questionnaire, the frequencies of febrile URI and episodes of severe exacerbations were 1.54 and 0.54 per subject per year, respectively. Logistic regression analysis showed that an older age [odds ratio (OR): 1.57; 95% confidence interval (CI): 1.15-2.13], female sex (OR: 1.58; 95% CI: 1.20-2.08), recurrent febrile episodes of URI (OR: 2.68; 95% CI: 1.47-4.91), a history of previous exacerbation within 1 year (OR: 1.74; 95% CI: 1.28-2.34), disuse of inhaled corticosteroids (ICSs) (OR: 2.63; 95% CI: 1.68-4.12), and disuse of add-on leukotriene receptor antagonists (LTRAs) (OR: 1.42; 95% CI: 1.06-1.74) were independently associated with moderate to severe symptom-severity. Poisson regression analysis showed that the independent factors that contributed to the frequency of recurrent severe exacerbations were female sex (regression coefficient ß = 0.62, p < .01), an episode of sputum with coughing (ß = 1.23, p < .01), nocturnal awakening (ß = 1.22, p < .01), and severe exacerbation (ß = 0.78, p < .01) within the previous 6 months. CONCLUSION: Symptom-severity of asthma and the frequency of severe exacerbations were associated with previous exacerbations and susceptibility to URI.

SOCS-1 participates in negative regulation of LPS responses.

SOCS-1 is a negative regulatory molecule of the JAK-STAT signal cascade. Here, we demonstrate that SOCS-1 is a critical downregulating factor for LPS signal pathways. SOCS-1 expression was promptly induced in macrophages upon LPS stimulation. SOCS-1-deficient mice were highly sensitive to LPS-induced shock and produced increased levels of inflammatory cytokines. Introduction of SOCS-1 inhibited LPS-induced NF-kappaB and STAT1 activation in macrophages. Furthermore, LPS tolerance, a refractory state to second LPS stimulation, was not observed in SOCS-1-deficient mice. These results suggest SOCS-1 as an essential, negative regulator in LPS responses that protects the host from harmful overresponses to LPS and may provide new insight into the endotoxin-induced fatal syndrome that occasionally occurs following infection.