기관지 천식환자에 있어 고장성 생리식염수 흡입을 이용한 유도객담 검사 - 검사법의 확립과 유도객담내 염증세포 관찰 -

강석민(Suck Min Kang),김철우(Chul Woo Kim),박중원(Joong Won Park),홍천수(Chun Soo Hong) 대한내과학회 1997 대한내과학회지 Vol.52 No.6

Objectives: From asymptomatic asthmatic patients unable to produce sputum spontaneously, we performed a non-invasive method obtaining sputum via inhalation of ultrsonically nebulised hyperosmolar saline(3%) which provides a way to investigate airway inflammation, And we also evaluated the effectiveness of the ultrasonic nebulizer in these patients. Methods: 50 asymptomatic patients unable to produce sputum spontaneously inhaled a hyperosmolar saline via an Ultrasonic nebulizer to induce sputum. We examined the quality of the sputum, total cell counts and differential cell counts of eosinophil after sputum processing. Results: 1) The quality of the sputum was uninterpretable (12%); poor(16%); fair(34%); good(38%), repectively. The mean induction time of expectoration was about 10 minutes. 2) The most common complaint was of a salty taste(45 cases). The mean fa11 in peak expiratory flow rate during inhalation of saline was 4.0% and the fall more than 20% was observed in 5 cases. 3) The mean eosinophil percent in induced sputum was 38.5%, The mean value of the total IgE and total peripheral eosinophil count was 818.2 IU/ml, 401.2/mm³ respectively, 4) There were no relationships between the sputum eosinophil count(%) and both the total IgE and the PC20(r=0.13, r=0.18). There was also an insignificant correlation between the sputum eosinophil count(%) and the total peripheral eosinophil count(r=D.22). Conclusion: We find that there are many eosinophils within induced sputum via inhalation of ultrsonically nebu1ised hyperosmolar saline from asymptomatic asthmatic patients unable to produce sputum spontaneously, Therefore we suggest that an induced sputum study via inhalation of hyperosmolar saline is useful to determine a patient`s status of airway inflammation.

폐결핵 진단에서 유도객담과 기관지경 검사의 유용성 비교

나문준(Moon Jun Na) 대한내과학회 1998 대한내과학회지 Vol.55 No.1

Objective: Expectorated sputum examination applied first in all patients with suspected pulmonary tuberculosis is low sensitivity. Bronchoscopy is applied next in patients with smear negative sputum. In some instances, bronchoscopy can not be available in patients because of poor conditions or cooperations. The aim of this study was to determine whether sputum induction is more simple and more convenient method than bronchoscopy or not to diagnose pulmonary tubereulosis in AFB smear-negative patients. Methods : The objects of this study were 47 patients whose smear results of expectorated sputum were negative or could not produce expectorated sputum, Consecutive patients underwent sputum induction with 3% hypertonic saline delivered by ultrasonic nebulizer at least Z4 hour before bronchial washing by fiber-optic bronchoscopy. All specimens were examined for acid-fast bacilli with Ziehl-Neelsen stain and cultured for mycobacteria. Results : 1) Sensitivity of AFB smear were 36% (17/47) in induced sputum, 53%(25/47) in bronchoscopy, but statistically not significant(p>0.05). 2) Sensitivity of mycobacterial culture were 53%(25/47) in induced sputum, 68%(32/47) in bronchoscopy, statistically not significant(p>0,05), 3) Positive AFB smear in endobronchial tuberculosis(n=9) were 5 in induced sputum, and 9 in bronchoscopy. 4) In 10 patients who were not obtainable sputum initially, AFB smear were positive in 7 in induced sputum and 9 in bronchoscopy 5) In ll patients who cultured in expectorated sputum later, positive culture were 10 patients both in induced sputum and bronchoscpy. 6) All 5 patients who didn't undergo bronchoscopy were positive AFB smear in induced sputum Conclusion : 1n suspected active pulmonary tuberculosis patients with negative AFB smear of expectorated sputum, induced sputum does not replace bronchoscopy to diagnose active pulmonary tuberculosis. But, sputum induction is useful diagnostic method to detect mycobacteria before or in not available bronchoscopy.

Comparative Evaluation of Three Homogenization Methods for Isolating Middle East Respiratory Syndrome Coronavirus Nucleic Acids From Sputum Samples for Real-Time Reverse Transcription PCR

성흥섭,용동은,기창석,김재석,성문우,이혁민,김미나 대한진단검사의학회 2016 Annals of Laboratory Medicine Vol.36 No.5

Background: Real-time reverse transcription PCR (rRT-PCR) of sputum samples is commonly used to diagnose Middle East respiratory syndrome coronavirus (MERS-CoV) infection. Owing to the difficulty of extracting RNA from sputum containing mucus, sputum homogenization is desirable prior to nucleic acid isolation. We determined optimal homogenization methods for isolating viral nucleic acids from sputum. Methods: We evaluated the following three sputum-homogenization methods: proteinase K and DNase I (PK-DNase) treatment, phosphate-buffered saline (PBS) treatment, and N-acetyl-L-cysteine and sodium citrate (NALC) treatment. Sputum samples were spiked with inactivated MERS-CoV culture isolates. RNA was extracted from pretreated, spiked samples using the easyMAG system (bioMérieux, France). Extracted RNAs were then subjected to rRT-PCR for MERS-CoV diagnosis (DiaPlex Q MERS-coronavirus, SolGent, Korea). Results: While analyzing 15 spiked sputum samples prepared in technical duplicate, false-negative results were obtained with five (16.7%) and four samples (13.3%), respectively, by using the PBS and NALC methods. The range of threshold cycle (Ct) values observed when detecting upE in sputum samples was 31.1-35.4 with the PK-DNase method, 34.7-39.0 with the PBS method, and 33.9-38.6 with the NALC method. Compared with the control, which were prepared by adding a one-tenth volume of 1:1,000 diluted viral culture to PBS solution, the ranges of Ct values obtained by the PBS and NALC methods differed significantly from the mean control Ct of 33.2 (both P<0.0001). Conclusions: The PK-DNase method is suitable for homogenizing sputum samples prior to RNA extraction.

Usefulness of Sputum Induction with Hypertonic Saline in a Real Clinical Practice for Bacteriological Yields of Active Pulmonary Tuberculosis

( Gil Myeong Seong ),( Jae Chun Lee ),( Jong Hoo Lee ),( Jeong Hong Kim ),( Mi Ok Kim ) 대한결핵 및 호흡기학회 2014 Tuberculosis and Respiratory Diseases Vol.76 No.4

Background: Mycobacterial identification in active pulmonary tuberculosis (APTB) is confirmative, even though successful rates using self-expectorated sputum are limited. Sputum specimens collected by hypertonic saline nebulization showed higher bacteriologic diagnostic sensitivities over those of self-expectoration, mostly studied in smear-negative or sputum-scarce patients. The efficacy of induced sputum was rarely assessed in real clinical settings. Methods: A prospective randomized case-control study was performed in one hospital. The subjects highly suspicious of APTB were asked to provide 3 pairs of sputum specimens in 3 consecutive days. The first pairs of the specimens were obtained either by self-expectoration (ES) from the next day of the visit or sputum induction with 7% saline nebulization in clinic (SI), and the other specimens were collected in the same way. The samples were tested in microscopy, culture, and polymerase chain reaction (PCR). The outcomes of the bacteriological diagnosis were compared. Results: Seventy six patients were assigned to either ES (38 subjects, median age of 51, 65.8% male) or SI (38 subjects, median age of 55, 52.6% male). APTB was clinically confirmed in 51 patients (70.8%), 27 in ES and 24 in SI. Among the APTB, more adequate specimens were collected from SI (41/65, 63.1%) than ES (34/80, 42.5%) (p=0.01). Bacteriological confirmation was achieved in 14 (58.3%) patients in SI, and 13 (48.1%) in ES (p=0.46). In the same-day bacteriological diagnosis with microscopy and PCR, there were positive results for 9 patients (37.5%) in SI and 7 patients (25.9%) in ES (p=0.37). Conclusion: Sputum induction improves sputum specimen adequacy. It may be useful for the same-day bacteriological diagnosis with microscopic examination and PCR.

The assessment of host and bacterial proteins in sputum from active pulmonary tuberculosis

Hsin-Chih Lai,Yu-Tze Horng,Pen-Fang Yeh,Jann-Yuan Wang,Chin-Chung Shu,Chia-Chen Lu,Jang-Jih Lu,Jen-Jyh Lee,Po-Chi Soo 한국미생물학회 2016 The journal of microbiology Vol.54 No.11

Pulmonary tuberculosis (TB) is caused by Mycobacterium tuberculosis. The protein composition of sputum may reflect the immune status of the lung. This study aimed to evaluate the protein profiles in spontaneous sputum samples from patients with active pulmonary TB. Sputum samples were collected from patients with pulmonary TB and healthy controls. Western blotting was used to analyze the amount of interleukin 10 (IL-10), interferon-gamma (IFN-γ), IL-25, IL- 17, perforin-1, urease, albumin, transferrin, lactoferrin, adenosine deaminase (also known as adenosine aminohydrolase, or ADA), ADA-2, granzyme B, granulysin, and caspase- 1 in sputum. Results of detection of IL-10, IFN-γ, perforin- 1, urease, ADA2, and caspase-1, showed relatively high specificity in distinguishing patients with TB from healthy controls, although sensitivities varied from 13.3% to 66.1%. By defining a positive result as the detection of any two proteins in sputum samples, combined use of transferrin and urease as markers increased sensitivity to 73.2% and specificity to 71.1%. Furthermore, we observed that the concentration of transferrin was proportional to the number of acidfast bacilli detected in sputum specimens. Detection of sputum transferrin and urease was highly associated with pulmonary TB infection. In addition, a high concentration of transferrin detected in sputum might correlate with active TB infection. This data on sputum proteins in patients with TB may aid in the development of biomarkers to assess the severity of pulmonary TB.

Usefulness of Sputum Induction with Hypertonic Saline in a Real Clinical Practice for Bacteriological Yields of Active Pulmonary Tuberculosis

성길명,이재천,이종후,김정홍,김미옥 대한결핵및호흡기학회 2014 Tuberculosis and Respiratory Diseases Vol.76 No.4

Background: Mycobacterial identification in active pulmonary tuberculosis (APTB) is confirmative, even though successful rates using self-expectorated sputum are limited. Sputum specimens collected by hypertonic saline nebulization showed higher bacteriologic diagnostic sensitivities over those of self-expectoration, mostly studied in smear-negative or sputum-scarce patients. The efficacy of induced sputum was rarely assessed in real clinical settings. Methods: A prospective randomized case-control study was performed in one hospital. The subjects highly suspicious of APTB were asked to provide 3 pairs of sputum specimens in 3 consecutive days. The first pairs of the specimens were obtained either by self-expectoration (ES) from the next day of the visit or sputum induction with 7% saline nebulization in clinic (SI), and the other specimens were collected in the same way. The samples were tested in microscopy, culture, and polymerase chain reaction (PCR). The outcomes of the bacteriological diagnosis were compared. Results: Seventy six patients were assigned to either ES (38 subjects, median age of 51, 65.8% male) or SI (38 subjects, median age of 55, 52.6% male). APTB was clinically confirmed in 51 patients (70.8%), 27 in ES and 24 in SI. Among the APTB, more adequate specimens were collected from SI (41/65, 63.1%) than ES (34/80, 42.5%) (p=0.01). Bacteriological confirmation was achieved in 14 (58.3%) patients in SI, and 13 (48.1%) in ES (p=0.46). In the same-day bacteriological diagnosis with microscopy and PCR, there were positive results for 9 patients (37.5%) in SI and 7 patients (25.9%) in ES (p=0.37). Conclusion: Sputum induction improves sputum specimen adequacy. It may be useful for the same-day bacteriological diagnosis with microscopic examination and PCR.

Efficacy of Induced Sputum for the Diagnosis of Pulmonary Tuberculosis in Adults Unable to Expectorate Sputum

( Jae Seuk Park ) 대한결핵 및 호흡기학회 2015 Tuberculosis and Respiratory Diseases Vol.78 No.3

Background: Induced sputum (IS) has been used to collect airway secretions in subjects who have inadequate sputum production. The aim of this study was to investigate the efficacy of IS for the diagnosis of pulmonary tuberculosis (PTB) in adults unable to expectorate sputum. Methods: Medical records of 39 PTB patients who underwent IS due to absence of spontaneous sputum production between January 2011 and March 2014 at a tertiary hospital in South Korea were reviewed. Results of acid fast bacilli smear, Mycobacterium tuberculosis culture and polymerase chain reaction assay for M. tuberculosis (TB-PCR) of IS specimens from these patients were analyzed. Clinical and high-resolution computed tomography (HRCT) characteristics were also analyzed to find characteristics associated with IS culture positivity. Results: Of the 39 IS specimens from PTB patients, 7 (17.9%) were smear positive and 31 (79.5%) were culture positive. Twenty-four IS specimens were tested for TB-PCR and 13 (54.2%) were positive on TB-PCR. Multivariate analysis showed that younger age (p=0.04) and presence of tree-in-bud appearance on HRCT (p=0.03) were independent predictors of IS culture positivity. Conclusion: IS is useful for the diagnosis of PTB in adults unable to expectorate sputum. Younger age and tree-in-bud appearance on HRCT were associated with IS culture positivity in these patients.

Usefulness of Sputum Induction with Hypertonic Saline in a Real Clinical Practice for Bacteriological Yields of Active Pulmonary Tuberculosis

Seong, Gil Myeong,Lee, Jaechun,Lee, Jong Hoo,Kim, Jeong Hong,Kim, Miok The Korean Academy of Tuberculosis and Respiratory 2014 Tuberculosis and Respiratory Diseases Vol.76 No.4

Background: Mycobacterial identification in active pulmonary tuberculosis (APTB) is confirmative, even though successful rates using self-expectorated sputum are limited. Sputum specimens collected by hypertonic saline nebulization showed higher bacteriologic diagnostic sensitivities over those of self-expectoration, mostly studied in smear-negative or sputum-scarce patients. The efficacy of induced sputum was rarely assessed in real clinical settings. Methods: A prospective randomized case-control study was performed in one hospital. The subjects highly suspicious of APTB were asked to provide 3 pairs of sputum specimens in 3 consecutive days. The first pairs of the specimens were obtained either by self-expectoration (ES) from the next day of the visit or sputum induction with 7% saline nebulization in clinic (SI), and the other specimens were collected in the same way. The samples were tested in microscopy, culture, and polymerase chain reaction (PCR). The outcomes of the bacteriological diagnosis were compared. Results: Seventy six patients were assigned to either ES (38 subjects, median age of 51, 65.8% male) or SI (38 subjects, median age of 55, 52.6% male). APTB was clinically confirmed in 51 patients (70.8%), 27 in ES and 24 in SI. Among the APTB, more adequate specimens were collected from SI (41/65, 63.1%) than ES (34/80, 42.5%) (p=0.01). Bacteriological confirmation was achieved in 14 (58.3%) patients in SI, and 13 (48.1%) in ES (p=0.46). In the same-day bacteriological diagnosis with microscopy and PCR, there were positive results for 9 patients (37.5%) in SI and 7 patients (25.9%) in ES (p=0.37). Conclusion: Sputum induction improves sputum specimen adequacy. It may be useful for the same-day bacteriological diagnosis with microscopic examination and PCR.

천식 의심 환자에서 천식 치료에 의한 유도 객담 내 염증지표의 변화

심명기 ( Myoung Ki Sim ),최인선 ( Inseon S. Choi ),김우진 ( Woo Jin Kim ) 대한천식알레르기학회 2012 천식 및 알레르기 Vol.32 No.3

Background: Eosinophilic airway inflammation is characteristic for typical asthma, but neutrophilic inflammation has been found in severe asthmatics resistant to steroid therapy. This study aimed to evaluate responses to asthma treatment according to inflammatory cell types in sputum. Methods: For a cross-sectional study, 240 consecutive patients with suspected asthma who underwent a 4.5% saline-induced sputum test and a test for airway hyperresponsiveness to methacholine or for airway reversibility were enrolled. Of these patients, 23 who inhaled corticosteroids were entered to a follow-up sputum study. Results: The number of sputum eosinophils was significantly related to the forced expiratory volume in one second/ forced vital capacity (r=-0.180, P<0.05) and airway hyperresponsiveness (r=-0.222, P<0.01), and the number of neutrophils was significantly related to age (r=0.165, P<0.05) and forced expiratory volume in one second (r=-0.157, P< 0.05). Subjects with sputum eosinophilia (n=88, 20.5%) had received asthma treatment before the tests more often than those with neutrophilia (n=23, 4.3%) or mixed granulocytic nature (n=11, 0%) (P<0.05 for each). In the follow-up study, the number of eosinophils was significantly decreased in the eosinophilic subjects (P<0.05), and the Δinterferon- γ/interleukin-5 ratio was significantly related to Δeosinophils (r=0.933, P<0.001). There was an inverse relationship between the number of neutrophils and the increase in forced expiratory volume in one second (r=-0.459, P<0.05) or Δinterferon-γ (r=-0.842, P<0.01). Conclusion: The severity of asthma may depend on inflammatory cell types in sputum. Although inflammatory markers of sputum improve by asthma treatment, treatment responses differ according to cell types. Therefore, sputum examination may be useful for predicting therapeutic responses in asthmatics.

Clinical Features of Eosinophilic Bronchitis

Kim, Yong-Hyun,Yun, Yeo-Il,Lim, Gune-Il,Park, Sang-Joon,Joo, Jae-Hak,Kim, Yong-Hon,Park, Choon-Sik 순천향대학교 2001 Journal of Soonchunhyang Medical Science Vol.7 No.2

Background : Eosinophilic inflammation of airway is usually associated with airway hyperresponsiveness in bronchial asthma. However, there is a small group of patients who has the eosinophilic inflammation in the bronchial tree with normal spirometry and no evidence of airway hyperresponsiveness, which was named eosinophilic bronchitis. The objectives of this study are 1) to investigate the incidence of eosinophilic bronchitis in the chronic cough syndrome and 2) to evaluate the clinical features and course of eosinophilic bronchitis. Methods : We evaluated 92 patients who had persistent cough for 3 weeks or longer In addition to usual diagnostic protocol, we performed differential cell count of sputum. Eosinophilic bronchitis was diagnosed when the patient had normal spirometric values, normal peak expiratory flow variability, no airway hyperresponsiveness, and sputum eosinophilia (>3%). Result : The causes of chronic cough were post-nasal drip in 33%, cough variant asthma in 16%, bronchitis in 15%, and eosinophilic bronchitis in 12% of the study subjects. Initial eosinophil percentage in sputum of patients with eosinophilic bronchitis was 26.8±6.1% (3.8-63.7%). Treatment with inhaled steroid is related with a subjective improvement of cough severity and a significant decrease of sputum eosinophil percentage (from 29.1±8.3% to 7.4±3.3%). During the follow up period of over ? months, recurrence of cough was associated with reappearance of sputum eosinophilia. Conclusion : Eosinophilic bronchitis is one of the important causes of chronic cough. Assessment of airway inflammation by sputum examination is important in investigating the cause of chronic cough, Cough in eosinophilic bronchitis are effectively controlled by inhaled corticosteroid, but it recurs frequently over a long term period of follow-up (Korean J Med 60:77-84, 2001) Key words : Bronchitis; Sputum; Eosinophilia