Isolation Of Tuberculosis Patients Research Articles

The Effectiveness of Chitosan and Snail Seromucous as Anti Tuberculosis Drugs

BACKGROUND: Tuberculosis (TB) disease is an infection caused by Mycobacterium tuberculosis (MTB) and is transmitted through sputum droplets of sufferers or suspect TB in the air. Chitosan has been widely used in the biomedical and pharmaceutical fields because it is a biocompatible, biodegradable, non-toxic, antimicrobial, and hydrating agent with positive effects on wound healing. Seromucous of snail has anti-tumor bioactivity and is non-toxic to lymphocyte cells, and can even stimulate lymphocyte proliferation. Seromucous of snail as glycoprotein containing carbohydrates; α-1 globulin-oromucoid fraction; glycans, peptides, glycopeptides, and chondroitin sulfate. AIM: This study was to determine the effectiveness of snail seromucous and chitosan as anti TB drugs (ATD) in vitro. METHODS: The research method is based on an experimental laboratory. MTB isolates in this research from sputum samples of patients suspected of TB in Surakarta Regional General Hospital. The stages of the study were performed MTB culture and identification, management sampling, and drug susceptibility testing. RESULTS: The research results showed chitosan 5%; a combination of chitosan 9% and snail seromucous 50% (ratio 1:1) is a microbicide against MTB TB patient isolates. Snail seromucous was ineffective as a microbicide against MTB TB patients. CONCLUSION: The effectiveness as a bactericide against MTB, chitosan, and its combination with snail seromucous has the potential to be an ATD alternative.

Revisiting the tuberculosis and leprosy sanatorium era: for the post COVID healthcare in India

Leprosy and tuberculosis (TB) are ancient diseases. These have been referred to in the Vedas and Ayurvedic Samhitas too. As no chemotherapy was effective against these diseases till middle of the 20th century, the main line of treatment was good food, open air and dry climate. Open air sanatoriums were formed for treatment and isolation of TB patients. The first sanatorium was founded in 1906 in Tiluania, near Ajmer, followed by one in Almora two years later. In 1909, the first non-missionary sanatorium was built near Shimla. Similarly, social stigma was the reason for creation of leprosy sanatoriums. First leprosy sanatorium was opened in 1925 at The school of tropical medicine, Calcutta and in 1955 the first research centre; The Lady Willingdon leprosy sanatorium, Chingleput (presently central leprosy teaching and research institute). These centers also contributed to knowledge of natural history of the disease and treatment. Subsequently many sanatoriums were set up at geographical locations which were at outskirts of cities, or at hilly terrains.1 One of the major reasons of setting up these sanatoria was to isolate the patients and prevent spread of disease further, in absence of medicines. Later on, many other infectious disease hospitals were also opened up, to cater to patients suffering from other infectious diseases.

Prevention of tuberculosis and isolation of tuberculosis patients in health care facilities

Prevention of tuberculosis and isolation of tuberculosis patients in health care facilities

Delayed isolation of smear-positive pulmonary tuberculosis patients in a Japanese acute care hospital

BackgroundActive pulmonary tuberculosis (TB) is associated with intra-hospital spread of the disease. Expeditious diagnosis and isolation are critical for infection control. However, factors that lead to delayed isolation of smear-positive pulmonary TB patients, especially among the elderly, have not been reported. The purpose of this study is to investigate factors associated with delay in the isolation of smear-positive TB patients.MethodsAll patients with smear-positive pulmonary TB admitted between January 2008 and December 2016 were included. The setting was a Japanese acute care teaching hospital. Following univariate analysis, significant factors in the model were analyzed using the multivariate Cox proportional hazard model.ResultsSixty-nine patients with mean age of 81 years were included. The median day to the isolation of pulmonary TB was 1 day with interquartile range, 1–4 days. On univariate analysis, the time to isolation was significantly delayed in male patients (p = 0.009), in patient who had prior treatment with newer quinolone antibiotics (p = 0.027), in patients who did not have chronic cough (p = 0.023), in patients who did not have appetite loss (p = 0.037), and in patients with non-cavitary lesion (p = 0.005), lesion located other than in the upper zone (p = 0.015), and non-disseminated lesion on the chest radiograph (p = 0.028). On multivariate analysis, the time to isolation was significantly delayed in male patients (hazard ratio [HR], 0.47; 95% confidence interval [CI], 0.25 to 0.89; P = 0.02), in patients who did not have chronic chough (HR, 0.52; 95% CI, 0.28 to 0.95; P = 0.033), and in patients with non-cavitary lesion on the chest radiograph (HR, 0.46; 95% CI, 0.23 to 0.92; P = 0.028).ConclusionsIn acute care hospitals of an aging society, prompt diagnosis and isolation of TB patients are important for the protection of other patients and healthcare providers. Delay in isolation is associated with male gender, absence of chronic cough, and presence of non-cavitary lesions on the chest radiograph.

Acquisition of second-line drug resistance and extensive drug resistance during recent transmission of Mycobacterium tuberculosis in rural China

Multidrug-resistant tuberculosis (MDR-TB) is prevalent in countries with a high TB burden, like China. As little is known about the emergence and spread of second-line drug (SLD) -resistant TB, we investigate the emergence and transmission of SLD-resistant Mycobacterium tuberculosis in rural China. In a multi-centre population-based study, we described the bacterial population structure and the transmission characteristics of SLD-resistant TB using Spoligotyping in combination with genotyping based on 24-locus MIRU-VNTR (mycobacterial interspersed repetitive unit-variable-number tandem repeat) plus four highly variable loci for the Beijing family, in four rural Chinese regions with diverse geographic and socio-demographic characteristics. Transmission networks among genotypically clustered patients were constructed using social network analysis. Of 1332 M. tuberculosis patient isolates recovered, the Beijing family represented 74.8% of all isolates and an association with MDR and simultaneous resistance between first-line drugs and SLDs. The genotyping analysis revealed that 189 isolates shared MIRU-VNTR patterns in 78 clusters with clustering rate and recent transmission rate of 14.2% and 8.3%, respectively. Fifty-three SLD-resistant isolates were observed in 31 clusters, 30 of which contained the strains with different drug susceptibility profiles and genetic mutations. In conjunction with molecular data, socio-network analysis indicated a key role of Central Township in the transmission across a highly interconnected network where SLD resistance accumulation occurred during transmission. SLD-resistant M. tuberculosis has been spreading in rural China with Beijing family being the dominant strains. Primary transmission of SLD-resistant strains in the population highlights the importance of routine drug susceptibility testing and effective anti-tuberculosis regimens for drug-resistant TB.

Risk factors for delayed isolation of tuberculosis patients in a tertiary care hospital in a low-incidence country

Results In the three-year time period, 58 patients had a cultureproven tuberculosis infection in our hospital, 20 with an extrapulmonary infection only. For 41 patients (70.7%), isolation measures were not taken. Twelve of these were patients with extrapulmonary tuberculosis only, and in these cases isolation measures were forgotten with aerosol generating procedures. The median number of days of non-isolation was 4 (range 1-162). For the patients with pulmonary tuberculosis (38 patients, 25 (65.8%) not isolated) four factors were predictive for non-isolation: presence of the symptom “fatigue”, absence of hemoptysis, absence of tuberculosis in the differential diagnosis, absence of involvement of a pulmonologist or infectious diseases specialist on admission. Due to these non-isolated patients, 1115 heath care workers and 78 patients had to be screened for tuberculosis.

Molecular epidemiology of Mycobacterium tuberculosis among South African gold miners.

HIV-associated tuberculosis remains a major health problem among the gold-mining workforce in South Africa. We postulate that high levels of recent transmission, indicated by strain clustering, are fueling the tuberculosis epidemic among gold miners. To combine molecular and epidemiologic data to describe Mycobacterium tuberculosis genetic diversity, estimate levels of transmission, and examine risk factors for clustering. We conducted a cross-sectional study of culture-positive M. tuberculosis isolates in 15 gold mine shafts across three provinces in South Africa. All isolates were subject IS6110-based restriction fragment length polymorphisms, and we performed spoligotyping analysis and combined it with basic demographic and clinical information. Of the 1,602 M. tuberculosis patient isolates, 1,240 (78%) had genotyping data available for analysis. A highly diverse bacillary population was identified, comprising a total of 730 discrete genotypes. Four genotypic families (Latin American Mediterranean spoligotype family; W-Beijing; AH or X; and T1-T4) accounted for over 50% of all strains. Overall, 45% (560/1,240) of strains were genotypically clustered. The minimum estimate for recent transmission (n - 1 method) was 32% (range, 27-34%). There were no individual-level risk factors for clustering, apart from borderline evidence for being non-South African and having self-reported HIV infection. The high M. tuberculosis genetic diversity and lack of risk factors for clustering are indicative of a universal risk for disease among gold miners and likely mixing with nonmining populations. Our results underscore the urgent need to intensify interventions to interrupt transmission across the entire gold-mining workforce in South Africa.

Characterisation of Mycobacterium tuberculosis isolates lacking IS6110 in Viet Nam

The molecular diagnosis of tuberculosis (TB) in Viet Nam is often based on the detection of insertion sequence (IS) 6110 in Mycobacterium tuberculosis. However, 8-11% of M. tuberculosis strains in South-East Asia do not contain this target and this undermines the validity of these molecular tests. We quantified the frequency of M. tuberculosis strains lacking IS6110 in rural Viet Nam and studied their epidemiological and clinical characteristics. Consecutively diagnosed adult TB patients in rural Southern Viet Nam submitted two sputum samples for culture, IS6110 restriction fragment length polymorphism (RFLP) spoligotyping and 15-loci variable number tandem repeat typing. Polymerase chain reaction (PCR) was performed to confirm the absence of IS6110 elements in strains lacking IS6110 hybridisation in RFLP. Among 2664 TB patient isolates examined, 109 (4.1%) had no IS6110 element. Compared to other strains, these no-copy strains were less often resistant to anti-tuberculosis drugs, particularly to streptomycin (adjusted OR 0.2, 95%CI 0.1-0.5), and showed significant geographic variation. No associations with TB history or demographic factors were found. Strains without the IS6110 target pose a problem in Viet Nam as regards false-negative molecular TB diagnosis in PCR. Compared to other strains circulating in Viet Nam, no-copy strains are more susceptible to anti-tuberculosis drugs.

Delayed Suspicion, Treatment and Isolation of Tuberculosis Patients in Pulmonology/Infectious Diseases and Non-Pulmonology/Infectious Diseases Wards

Background/PurposeDelayed diagnosis and isolation increases the risk of nosocomial transmission of tuberculosis (TB). To assess the risk of delayed management of TB, we analyzed the risk factors of prolonged delay in isolation of smear-positive TB patients in pulmonology/infectious diseases and other wards in a tertiary teaching hospital.MethodsWe enrolled smear-positive TB patients aged > 16 years with delayed respiratory isolation following hospitalization. Medical records were reviewed retrospectively. Time intervals between admission, order of sputum acid-fast staining, initiation of anti-tuberculous treatment and isolation were compared between pulmonology/infectious diseases wards (PIWs) and other wards. Risk factors were analyzed in patients with prolonged isolation delay of > 7 days in individual groups.ResultsIsolation was delayed in 191 (73.7%) of 259 hospitalized smear-positive TB patients. Median suspicion, treatment and isolation delays were 0, 3 and 4 days in PIWs and 1, 5 and 7 days in other wards. For patients admitted to non-PIWs, atypical chest radiographs, symptoms without dyspnea or not being admitted from the emergency department (ED) were risk factors for prolonged isolation delay exceeding 7 days. The only risk factor for delayed isolation in patients admitted to PIWs was age ≥ 70 years.ConclusionDelays in suspicion, treatment and isolation of TB patients were longer in non-PIWs. Clinicians should be alert to those admitted to non-PIWs with atypical chest radiographs, atypical symptoms, or not admitted from the ED.

Genomic interrogation of ancestral Mycobacterium tuberculosis from south India

Mycobacterium tuberculosis is a very important global pathogen. One quarter of the world's TB cases occur in India. The tuberculosis strains isolated from south Indian patients exhibit certain phenotypic characteristics like low virulence in guinea-pigs, resistance to isoniazid, thiophene-2-carboxylic acid hydrazide (TCH) and para-amino salicylic acid (PAS), and enhanced susceptibility to H 2O 2. Besides this, a large percentage of the isolates harbor only a single copy of IS 6110 which makes these strains distinct. Hence, we have studied the genotypic characteristics of these strains by using advanced techniques like Deletion Micro array, deletion PCR, allelic discrimination RT-PCR using several lineage specific markers and KatG G1388T (non-synonymous) polymorphism along with spoligotyping. The analysis of 1215 tuberculosis patient isolates from south India revealed that 85.2% belonged to the ancestral lineage of M. tuberculosis. Comparative whole-genome hybridization identified six new genomic regions within this lineage that were variably deleted.

Detection of rpoB Mutations Associated with Rifampin Resistance in Mycobacterium tuberculosis Using Denaturing Gradient Gel Electrophoresis

Denaturing gradient gel electrophoresis (DGGE) was used to probe for mutations associated with rifampin (RIF) resistance in the rpoB gene of Mycobacterium tuberculosis. DGGE scans for mutations across large regions of DNA and is comparable to DNA sequencing in detecting DNA alterations. Specific mutations are often recognized by their characteristic denaturation pattern, which serves as a molecular fingerprint. Five DGGE primer sets that scanned for DNA alterations across 775 bp of rpoB were developed. These primer sets were used to scan rpoB for DNA alterations in 296 M. tuberculosis patient isolates from the United States-Mexico border states of Texas and Tamaulipas. The most useful primer set scanned for mutations in the rifampin resistance-determining region (RRDR) and detected mutations in 95% of the RIF-resistant isolates compared to 2% of RIF-susceptible isolates. Thirty-four different alterations were observed within the RRDR by DGGE. In addition, isolates harboring mixtures of DNA within rpoB were readily detected by DGGE. A second PCR primer set was used to detect the V146A mutation in 5 to 7% of RIF-resistant isolates. A third primer set was used to detect mutations in 3% of RIF-resistant isolates, some of which also harbored mutations in the RRDR. Only 1 of 153 RIF-resistant isolates did not have a detectable rpoB mutation as determined by DGGE and DNA sequencing. These results demonstrate the power and usefulness of DGGE in detecting mutations associated with drug resistance in M. tuberculosis.

Attitude and social consequences of tuberculosis in Addis Ababa, Ethiopia.

To gain a better understanding of the attitude and social consequences of tuberculosis (TB) in Addis Ababa, Ethiopia. A cross sectional survey using a structured questionnaire and a qualitative study based on the focus group discussion (FGD) technique. Eight different kebeles (urban dwellers' associations) of six woredas (next higher administrative level to kebele). Seven hundred and three participants, comprising 326 males and 377 females were interviewed using the written questionnaire and 36 recent and current TB patients through focus group discussion. Data were analysed using SPSS/PC statistical package. The proportions were compared using univariate and bivariate analyses to show the frequency distribution and evaluate the relationship among different variables. In FGD, topics relevant to the research questions were identified, sorted and analysed. Conclusions were then formulated. Attitude of participants towards TB and its victims and social consequences of being a TB patient. Eighty three per cent of the respondents were aware that TB is a disease transmitted from one person to another and 80.1% perceived TB as an extremely severe disease. Of the total, 81.5% answered that TB is caused by cold, 69.0% feel that TB patients are not accepted in the community and 78.3% fear physical contact with TB patients. Most participants of the FGD sessions agreed that TB is a very dangerous, contagious but curable disease and the community has a generally negative attitude towards them. They also think that TB is associated with HIV/AIDS in the society. These attitudes have social consequences particularly the stigmatization and social isolation of TB patients. Four hundred and four (57.5%) respondents were found to be in favour of the short course chemotherapy against 226 (32.1%) choosing the longer course. Clinical improvement and unavailability of some drugs or their cost were the first and second main reasons indicated, respectively, for defaulting by respondents. The perception by most respondents that tuberculosis is incurable, transmittable and associated with HIV/AIDS, led to the understanding that TB is a very dangerous disease. This, in turn, contributes to social avoidance and the resultant consequences in TB patients. Health education must be stepped-up within the TB control programme, and the psychosocial implications of TB should be given due attention.

Genetic diversity of Mycobacterium tuberculosis in Sicily based on spoligotyping and variable number of tandem DNA repeats and comparison with a spoligotyping database for population-based analysis.

In a previous study, we proposed to associate spoligotyping and typing with the variable number of tandem DNA repeats (VNTR) as an alternative strategy to IS6110-restriction fragment length polymorphism (RFLP) for molecular epidemiological studies on tuberculosis. The aim of the present study was to further evaluate this PCR-based typing strategy and to describe the population structure of Mycobacterium tuberculosis in another insular setting, Sicily. A collection of 106 DNA samples from M. tuberculosis patient isolates was characterized by spoligotyping and VNTR typing. All isolates were independently genotyped by the standard IS6110-RFLP method, and clustering results between the three methods were compared. The totals for the clustered isolates were, respectively, 15, 60, and 82% by IS6110-RFLP, spoligotyping, and VNTR typing. The most frequent spoligotype included type 42 that missed spacers 21 to 24 and spacers 33 to 36 and derived types 33, 213, and 273 that, together represented as much as 26% of all isolates, whereas the Haarlem clade of strains (types 47 and 50, VNTR allele 32333) accounted for 9% of the total strains. The combination of spoligotyping and VNTR typing results reduced the number of clusters to 43% but remained superior to the level of IS6110-RFLP clustering (ca. 15%). All but one IS6110-defined cluster were identified by the combination of spoligotyping and VNTR clustering results, whereas 9 of 15 spoligotyping-defined clusters could be further subdivided by IS6110-RFLP. Reinterpretation of previous IS6110-RFLP results in the light of spoligotyping-VNTR typing results allowed us to detect an additional cluster that was previously missed. Although less discriminative than IS6110-RFLP, our results suggest that the use of the combination of spoligotyping and VNTR typing is a good screening strategy for detecting epidemiological links for the study of tuberculosis epidemiology at the molecular level.

Different tuberculosis in men and women: beliefs from focus groups in Vietnam

After decades in decline, tuberculosis (TB) has been increasing worldwide. In 1993, the World Health Organisation declared TB a global emergency. Passive case-finding is an important part of TB control programmes, and this is strongly affected by people’s perceptions and beliefs of TB and society’s behaviour towards TB sufferers. The aim of this study was to describe the perceptions and beliefs of Vietnamese people regarding TB and its risk factors with special reference to differences between men and women. Sixteen focus group discussions (FGDs) were organised in four districts representing different regions in Vietnam and consisting of men and women, TB patients and non-TB participants. In general, participants had good knowledge of TB being a dangerous, contagious and infectious disease, caused by germs. However, traditional beliefs in different types of TB still exist, mainly among older people in rural areas, but also resorted to by other people once ill. Four main types of TB were reported: (1) ‘ Lao truyen’ (hereditary TB), handed down from older generations to latter ones through ‘family blood’, regardless of sexes; (2) ‘ Lao luc’ (physical TB), caused by hard work, more men affected; (3) ‘ Lao tam’ (mental TB), caused by too much worrying—more women affected; and (4) ‘ Lao phoi’ (lung TB), dangerous and caused by TB germs, transmitted through the respiratory system—more men affected. Other general risk factors were also mentioned. Men were perceived to get TB more often than women, as they were more exposed to risk factors during both work and leisure time. These traditional beliefs may contribute to long delays to TB diagnosis and increased social stigma and isolation of TB patients and their families due to erroneous beliefs in transmission routes. Our findings demonstrate areas where TB control programmes may be improved.

Enhanced capacity of a widespread strain of Mycobacterium tuberculosis to grow in human macrophages.

To determine whether the extent of spread of Mycobacterium tuberculosis strains in the community correlated with their capacity to replicate in human macrophages, intracellular growth rates of M. tuberculosis patient isolates were measured. Strain 210 caused disease in 43 patients in central Los Angeles, 3 "small-cluster" strains caused disease in 8-23 patients, and 5 "unique" strains each caused disease in only 1 patient who was positive by sputum acid-fast smear and spent substantial amounts of time at homeless shelters that were tuberculosis transmission sites. Strain 210 isolates grew significantly more rapidly than small-cluster and unique strains in macrophages. All strains elicited production of similar amounts of tumor necrosis factor-alpha, interleukin (IL)-6, IL-10, and IL-12 and were equally susceptible to reactive nitrogen intermediates. It was concluded that the extensive spread of an M. tuberculosis strain correlated with its capacity to replicate rapidly in human macrophages, which may be a marker of virulence.

Respiratory isolation of tuberculosis patients using clinical guidelines and an automated clinical decision support system.

To evaluate a clinical guideline and an automated computer protocol for detection and respiratory isolation of tuberculosis (TB) patients. An automated computer protocol was tested on a retrospective cohort of adult culture-positive TB patients admitted from 1992 to 1993 to Columbia-Presbyterian Medical Center and evaluated prospectively from July 1995 until July 1996. A large teaching hospital in New York City. 171 adult patients admitted from 1992 to 1993 and 43 patients admitted between July 1995 and July 1996. The 1990 Centers for Disease Control and Prevention guidelines for preventing transmission of TB were adapted to formulate clinical guidelines to ensure early isolation of TB patients at Columbia-Presbyterian Medical Center. Implementation of a clinical respiratory isolation protocol resulted in a significant improvement in TB patient isolation rates, from 45 (51%) of 88 in 1992 to 62 (75%) of 83 in 1993 (P<.001). In testing automated protocols, the theoretical improvement would have identified an additional 27 patients not isolated by clinicians, making the overall isolation rate 134 (78%) of 171. For the prospective evaluation, 30 (70%) of 43 TB patients were isolated by clinicians adhering to the clinical protocol. Four additional patients were identified by the automated TB protocol, making the combined isolation rate 34 (79%) of 43. A clinical policy to isolate TB patients and suspected human immunodeficiency virus-infected patients with cough, fever, or radiographic abnormalities improved isolation of culture-documented TB patients from 1992 to 1993. Automated computer protocols were successful in identifying additional potentially infectious patients that clinicians failed to place on respiratory isolation. Clinical and automated protocols combined resulted in better isolation rates than a clinical protocol alone.

Respiratory Isolation of Tuberculosis Patients Using Clinical Guidelines and an Automated Clinical Decision Support System

Respiratory Isolation of Tuberculosis Patients Using Clinical Guidelines and an Automated Clinical Decision Support System

Studies of polymorphic DNA fingerprinting and lipid pattern of Mycobacterium tuberculosis patient isolates in Japan.

Strain differentiation by DNA restriction fragment length polymorphism (RFLP) has been used mainly for the epidemiological purpose of Mycobacterium tuberculosis infection. In this study, we tried to connect the molecular and phenotypic characteristics of M. tuberculosis patient isolates by comparing the DNA fingerprints obtained by RFLP using IS6110 and lipid patterns using two-dimensional thin-layer chromatography (2-D TLC) with silica gel, since M. tuberculosis has a lipid-rich cell envelope which contributes to the virulence and immunomodulatory properties. We found that 66 isolates of M. tuberculosis from tuberculosis patients showed that the occurrence of IS6110 varied from 1 to 24 copies. The IS6110 patterns were highly variable among isolates. Fifty different RFLP patterns were observed, and 12 RFLP patterns were shared by two or more strains. By computerized analysis of the RFLP patterns of M. tuberculosis patient isolates, we found that 95% of the isolates fell into seven clusters, from A to G, with at least two isolates in each (> 30% similarity). Among the cellular lipids, the phospholipid composition did not differ by strain, whereas the glycolipid pattern differed markedly. Especially, the relative concentration of cord factor and sulfolipid, both of which were known as virulent factors, varied by strain. The fingerprints of some strains showed an association between the DNA and glycolipid patterns, even though some of the same DNA fingerprint strains showed differences in lipid patterns. Among the patient isolates, M. tuberculosis strain 249 possessed a specific glycolipid with 2-O-methyl-L-rhamnose and L-rhamnose, which is rarely found in other strains. This glycolipid showed serological activity against the sera of tuberculosis patients, even if the reactivity was not as strong as trehalose dimycolate. It also showed the inhibition of phagosome-lysosome fusion in macrophages, suggesting involvement with virulence. These results suggest that RFLP analysis using IS6110 is useful for clustering the human isolates of M. tuberculosis, however, for further strain differentiation on virulence, a lipid analysis provides more information.

Evaluation of sputum smears concentrated by cytocentrifugation for detection of acid-fast bacilli

Early identification and isolation of tuberculosis patients is of utmost importance to minimize the risk of further epidemic spread of the disease. The traditional concentrated acid-fast smears are not very reliable tools for the presumptive diagnosis of tuberculosis. Acid-fast bacillus (AFB) smears from 120 patients specimens and 80 simulated AFB samples were processed according to standard laboratory procedures and by cytocentrifugation (Cyto-Tek, Ames Division, Miles Laboratories, Inc., Elkhart, Ind.). Prior to dispensing of samples into the Cyto-Tek chambers, specimens were liquefied and decontaminated by mixture with an equal volume of 5% sodium hypochlorite (household bleach). Culture and smear results were correlated. Of 120 patient specimens, 43 were culture and smear negative by both methods. Ten specimens were overgrown with mold and bacteria, but 2 of them had positive AFB smears by cytocentrifugation only. There were 67 positive AFB cultures, with 67 positive cytocentrifuge smears and 34 positive smears by the conventional technique. Of the 80 simulated positive AFB samples, all grew mycobacteria on culture. Smears from the 10(5)- to 10(3)-CFU/ml specimens were positive by both methods. The simulated samples with 10(2) CFU/ml yielded smears positive only by cytocentrifugation. The Cyto-Tek AFB smears had a greater correlation with positive culture than did the smears from concentrated specimens. The sensitivity, efficiency, and rapidly of the Cyto-Tek AFB smear technique resulted in increased detection of mycobacteria in clinical specimens. The simplicity and safety of this method will enable qualified mycobacteriology technologists to rapidly and accurately perform sputum smears for AFB at clinics, emergency rooms, and field sites, as well as in the traditional laboratory setting.