Rapid dot sputum and serum assay in pulmonary tuberculosis

A rapid direct sputum (Sp.) and/or antibody assay, based on immunoblotting and enzyme immunoassay is described. The test can detect mycobacterial antigens or antibodies in clinical specimens from pulmonary tuberculosis (TB) patients. In this study, 87 sputa, 87 sera and 40 paired sputa and sera were utilized from smear-positive and smear-negative, culture-positive patients; 59 sputa, 37 sera and 22 paired sputa and sera from nontuberculosis respiratory disease patients and 68 sera from healthy controls. The antigen detection in sputum by dot assay has 86.1% sensitivity on active tuberculosis patients, 92.9% specificity, 91.6% positive predictive value (PPV), 88.2% negative predictive value (NPV) and 10.3% error. The antibody assay has 83.6% sensitivity, 95.4% specificity, 94.4% positive predictive value, 85.6% negative predictive value and 11% error. The test performed on paired sputum and serum (Sr.) samples has a sensitivity of 93.3%, which rose to 96.1% on smear-positive and culture-positive patients, but the specificity decreased to 83% in sputum, whereas in serum it was 92%. The results of the assay, combined with clinical and radiological data, could form the basis for starting an earlier course of treatment for tuberculosis.     

A mite survey in the dumpsters for urban solid rubbish

Summary The presence of house dust mites and storage mites in dumpsters was investigated in 3 different areas of Venice hinterland. The aim of this study was to find a relationship between some reported respiratory symptoms among 6 workers who were responsible for moving the dumpsters to the truck and the bio-aerosol released from such operation. These symptoms were closely related to the work and after allergological evaluation we found sensitisation to house dust mites in all 6 employees. The mine exposure in the workplace was assessed by a standardized sampling and analysis of dust obtained after brushing the inside surfaces of dumpsters. In 50% of the specimens (10/20) we found mites belonging to the following families: Pyroglyphidae (Dermatophagoides spp.), Acaridae (Acarus siro) and a lower amount of Cheyletidae (Cheyletus spp.). An indirect test (guanine test) was also performed and gave positivity in 16 dust samples (80%). The insides of dumpsters showed to be an optimal environment for mites survival and reproduction because of cracks and grooves on the walls, the high humidity level and the presence of mould. These mites probably come from dwelling places and other environments (markets, slaughters-houses, farms, etc.). This suggests that mites allergens can be released during rubbish discharging from the dumpsters and represent a possible risk for the employees.     

A Comparative Study of Mycobacteria from Patients' Room Dusts and from Sputa of Tuberculous Patients

The source of mycobacteria other than Mycobacterium tuberculosis occurring in sputa of tuberculous patients as casual isolates was investigated by comparing the occurrence rate of mycobacterial species between patient's room dust and patient's sputum. Almost all species of mycobacteria recovered from sputa could be found in dusts obtained from the rooms. However, the percentage of occurrence of the mycobacterial species in dusts differed from that in sputa. In dusts, Mycobacterium fortuitum (39.6%), Mycobacterium nonchromogenicum (23.5%) and Mycobacterium gordonae (16.7%) occurred in high frequencies, whereas Mycobacterium intracellulare (69.6%), M. gordonae (5.9%), Mycobacterium scrofulaceum (5.2%), and Gordona bronchialis (10.4%) were the main species found in sputa. The patterns of occurrence of mycobacterial species as illustrated above may suggest that pathogenic mycobacteria survive in the respiratory tract while nonpathogenic ones are destroyed there, thus the human body acts as a selective medium for the pathogenic mycobacterial species. Serotype studies on strains of M. intracellular as casual isolates indicated that those from sputa of tuberculous patients were different from those derived from patients with lung diseases due to this species of mycobacteria. These results led us to the conclusion that mycobacteria in patients' room dusts were the source of mycobacteria occurring in sputa as casual isolates, particularly the pathogenic mycobacteria in dusts are more likely to survive in the human respiratory tract, occasionally multiplying and causing disease under favorable circumstances.     

Comparison of conventional and molecular methods for the detection of bacterial pathogens in sputum samples from cystic fibrosis patients

The nature of the micro-flora present in sputa of six different cystic fibrosis (CF) patients was assessed using routine microbiological culture and molecular methods. Bacterial genes for the small subunit ribosomal RNA (ssu rDNA) were specifically amplified from DNA extracted from the sputum samples, cloned and characterised by hybridisation and DNA sequencing. A large number of clones from six sputa were screened. Initially, oligonucleotide hybridisation was performed with five probes, specific for Gram-positives and Gram-negatives in general and the main pathogens for the CF patient (Staphylococcus aureus, Pseudomonas aeruginosa and Haemophilus influenzae). For a single sputum sample, the results were fully congruent when culture and molecular methods were compared. In the other five sputa, discrepancies for S. aureus and/or H. influenzae were documented. Although S. aureus DNA and H. influenzae DNA was detected in three and four sputa, respectively, strains could not be cultured. Although the PCR approach is not capable of distinguishing viable from dead bacteria, all of the CF patients had a history of S. aureus infections, while one of the CF patients once had cultivable H. influenzae in the sputum as well. A number of clones for probe-unidentified Gram-negative or Gram-positive bacterial species were further analysed by sequencing and additional potential pathogens were identified. Although routine culture of sputum frequently points to mono-specific exacerbations, our molecular data indicate that the other CF-related pathogens appear to be persistently present as well. We conclude that routine culture for bacterial pathogens from CF sputa yields limited microbiological information since it frequently fails to identify a number of pathogenic bacterial species that are potentially present in a viable status in the lungs of these patients.     

The effect of sub-floor heating on house-dust-mite populations on floors and in furniture

It is well known that dehydrating conditions for house dust mites can be created by simply raising the temperature, causing loss of body water and eventually death. Thus, it can be expected that conditions for dust mites are less favourable on floors supplied with sub-floor heating. This was examined in a study of 16 houses with sub-floor heating and 21 without. The pattern of changes in air humidity and temperature on the floors was investigated and compared to known data of the tolerance of dust mites. Also the resident mite populations were compared. Floors with sub-floor heating had, on average, fewer mites, but the difference with unheated floors was small. It was remarkable that mite numbers were also lower in upholstered furniture. Another important observation was that some houses with sub-floor heating had high mite numbers, indicating that this type of heating is compatible with a thriving mite population. Temperature and humidity conditions of heated floors may allow mites not only to survive, but also to remain active in winter. A moderate increase in temperature, a moderate decrease inf (absolute) air humidity, or a combination of both, will suffice to keep the humidity all winter below the Critical Equilibrium Humidity, the level of air humidity that is critical for mite growth and reproduction, hence for allergen production. However, it is argued that measures to suppress allergen production by house dust mites are likely to be far more effective if taken in summer rather than in winter. This revised version was published online in July 2006 with corrections to the Cover Date.     

Distribution and abundance of house-dust mites (Acarina: Pyroglycphidae) in Rome,Italy

For the first time in Rome, house-dust mite infestation was studied in 90 randomly selected houses. In each house, mite infestation was assessed in three sites: mattress, bedroom and living room. In total, 87.8% of the sampled houses were positive for dust mites. In the houses infested, 11.4% showed densities of >100 mites/g of dust, 15.2% registered densities between 50 and 99, and in the remaining houses (73.4%), the densities were between 1 and 49 mites/g dust. The percentages of infested houses were positively correlated with the relative humidity (RH) values (r=0.89,P=0.02). At the lowest range of RH (between 46 and 50), the infestation was 50% and at the highest range of RH (between 73 and 78) it was 100%. The mattress was significantly the most infested (71.1%) of the tested sites. Only wool and spring mattresses were infested, and they did not show any significant differences in mite concentrations.Dermatophagoides farinae was the most abundant species (53.1%), followed byGlycyphagus domesticus (34.5%),D. pteronyssinus (5.2%), andEuroglyphus maynei (0.2%);D. farinae was also the most frequent species (56.9%). The remaining specimens (7.0%) were predator species commonly found in houses. The prevalence ofD. farinae in Rome is discussed.     

House-dust mite sensitivity among rural and urban asthmatics of West Bengal, India: a comparison

House-dust mite allergy is a fairly common problem in West Bengal among individuals sensitive to dust inhalation. House-dust mites belonging to the genusDermatophagoides are abundant in the homes of asthmatic patients residing in urban as well as rural areas of West Bengal. The frequency of positive skin reaction to different dust-related allergens tested was higher (χ²=5.4777, df = 1;P < 0.05) among patients of urban areas compared with that among the patients of rural areas. Urban patients showed more frequent skin reaction towards cockroach allergen, while rural patients are more sensitive to hay-dust allergen and these are very much related to their local environmental conditions. Analysis of radioallergosorbent test (RAST) results against house dust (HD) and mites reveal that 73 and 90% patients of both urban and rural areas responded positively towardsDermatophagoides pteronyssinus (DP) andDermatophagoides farinae (DF) antigens, respectively. The present study indicates no significant difference in house-dust mite sensitivity and mite levels in homes among the rural and urban asthmatics of West Bengal, India as evidenced from the results of analysis of dust samples, allergy skin test and detection of mite-specific IgE antibodies by RAST.     

Guanine as a hygienic index for allergologically relevant mite infestations in mattress dust

Since guanine is not only an essential constituent of vital nucleic acids, but also the main end product of nitrogenous waste excretion in arachnids, it is a potential candidate for a hygienic index for mite activity in house dust. The public health significance of these mites is based on their production of allergen in the home environment.In 101 mattress-dust samples, total number of mites and guanine content were positively correlated. The hygienic standard of 10 mites per 0.1 g dust (as derived from Danish investigations) was met in all samples with less than 0.06% guanine in the dry dust. Where the guanine content was more than or equal to 0.25% in the dry dust, mite numbers were higher than 10 mites per 0.1 g dust in 43 of the 44 samples. The three different groups (negligible mite content, possibly allergologically relevant mite concentration and allergologically unhygienic mite situation) could be easily discriminated on the color scale.The guanine-AZO-dye reaction in vacuum cleaner dust is a strong and promising candidate for an allergologically relevant hygienic index in the home.     

Isolation and characterization of microparticles in sputum from cystic fibrosis patients

Background

Microparticles (MPs) are membrane vesicles released during cell activation and apoptosis. MPs have different biological effects depending on the cell from they originate. Cystic fibrosis (CF) lung disease is characterized by massive neutrophil granulocyte influx in the airways, their activation and eventually apoptosis. We investigated on the presence and phenotype of MPs in the sputum, a rich non-invasive source of inflammation biomarkers, of acute and stable CF adult patients.

Methods

Spontaneous sputum, obtained from 21 CF patients (10 acute and 11 stable) and 7 patients with primary ciliary dyskinesia (PCD), was liquefied with Sputasol. MPs were counted, visualized by electron microscopy, and identified in the supernatants of treated sputum by cytofluorimetry and immunolabelling for leukocyte (CD11a), granulocyte (CD66b), and monocyte-macrophage (CD11b) antigens.

Results

Electron microscopy revealed that sputum MPs were in the 100-500 nm range and did not contain bacteria, confirming microbiological tests. CF sputa contained higher number of MPs in comparison with PCD sputa. Levels of CD11a⁺-and CD66b⁺-, but not CD11b⁺-MPs were significantly higher in CF than in PCD, without differences between acute and stable patients.

Conclusions

In summary, MPs are detectable in sputa obtained from CF patients and are predominantly of granulocyte origin. This novel isolation method for MPs from sputum opens a new opportunity for the study of lung pathology in CF.     

Efficacy and pattern of use of sputum cytology as a diagnostic test

Sputum cytology is regarded by many clinicians as a noninvasive, cheap and simple test for the diagnosis of bronchogenic carcinoma. Since the introduction of fibre-optic bronchoscopy and more easily obtained bronchial biopsies reliance on sputum cytology has diminished. However, in Edinburgh it was perceived that sputum samples were still being sent as well as, rather than instead of, bronchoscopic specimens. This retrospective study was undertaken to determine whether or not cytological examination of sputum is an efficient and sensitive test in the investigation of patients with suspected bronchogenic carcinoma. It demonstrated that the Lothian University Hospitals NHS Trust Pathology Directorate receives many sputa from departments not specializing in respiratory disease when there is no indication for the test. In addition, we have shown that the absolute sensitivity of the test is only 5% and that when there is a strong clinical suspicion of bronchogenic carcinoma the results of sputum cytology do not play a significant role in the management of the patient. We recommend that sputum cytology is restricted to those patients under the care of Respiratory Units in whom bronchoscopy is inappropriate or unsuccessful.     

Diagnostic Accuracy of the PURE-LAMP Test for Pulmonary Tuberculosis at the County-Level Laboratory in China

Background

Early and effective detection of Mycobacterium tuberculosis (MTB), particularly in smear-negative tuberculosis (TB), is a priority for global TB control. Loop-mediated isothermal amplification with a procedure for ultra rapid DNA extraction (PURE-LAMP) can detect TB in sputum samples rapidly and with high sensitivity and specificity. However, the PURE-LAMP test has not been effectively evaluated, especially in resource-limited laboratories. In this study, we evaluated the performance of the PURE-LAMP test for TB detection in TB suspects from two county-level TB dispensaries in China.

Methodology/Principal Findings

From April 2011 to February 2012, patients with suspected TB were continuously enrolled from two county-level TB laboratories in China. Three sputum samples (spot, night, and morning sputum) were collected from each recruited patient. Detection of MTB by PURE-LAMP was compared to a reference standard L-J culture. The results showed that the sensitivity of the PURE-LAMP test based on spot sputum for MTB detection was 70.67%, while the sensitivity of the PURE-LAMP test based on spot sputum for MTB detection in smear positive and culture positive patients and smear negative and culture positive patients was 92.12% and 53.81%, respectively. The specificity of PURE-LAMP based on spot sputum for MTB detection was 98.32%. The sensitivity and specificity of the PURE-LAMP test based on three sputa combination for MTB detection was 88.80% and 96.86%, respectively. The results also showed that the PURE-LAMP test had a significantly lower contamination rate than did solid culture.

Conclusions/Significance

The study suggested that, in peripheral-level TB laboratories in China, the PURE-LAMP test showed high sensitivity and specificity for TB detection in TB suspects, making it a more effective, rapid, and safe method worthy of broader use in the future.     

The Diagnosis of Pneumococcal Chest Infection by Counter-Current-Immunoelectrophoresis

A simple, inexpensive method of counter-current-immunoelectrophoresis was used to detect pneumococcal antigens in sputum, serum and pleural fluid. Antigens were detected in sputa from 69% of subjects with pneumococcal pneumonia and there was a good correlation with the certainty of the clinical diagnosis. Serum antigens were found in 12.5% of subjects with either a definite or a presumed diagnosis and in one case from a patient who was not thought to have pneumococcal pneumonia. The pathogenic importance of antigens in the sputa of patients with a history of chronic respiratory disease was uncertain. Counter-current-immunoelectrophoresis was more sensitive than routine bacteriological methods for demonstrating the presence of pneumococci in pleural fluids.     

Clinical Significance of Skin Reactions to Mite Extracts in Children with Asthma

The mite Dermatophagoides pteronyssinus has been identified in dust from the houses of children in Birmingham suffering from asthma. Skin tests were carried out on 150 asthmatic children with extracts of D. pteronyssinus, of the related species D. farinae, of other mites found in house dust, and of crude house dust. Though positive reactions to D. pteronyssinus were obtained more frequently and were of greater size than those to the other extracts, it was considered that D. farinae is a suitable substitute for D. pteronyssinus for skin testing.In further tests on 302 asthmatic children with mite extracts and with extracts of allergens obtained commercially reactions to the former extracts were much more common than reactions to the latter.Major skin reactions (weals with diameter of 5 mm. or more) were present in 77% of children with a history of perennial asthma and house-dust sensitivity. Hence allergy to house-dust mites, particularly D. pteronyssinus, is of considerable importance in childhood asthma, and further study of the ecology and control of the mites in dust is desirable.     

盐城地区未成年过敏性疾病患者吸入性过敏原IgE检测结果分析

摘要 目的：通过研究分析盐城地区未成年过敏性疾病患者吸入性过敏原特异性IgE检测结果分布变化特点,为过敏性疾病预防和临床诊疗提供科学依据。方法：自2020年1月至2021年3月期间,选择430例未成年(<18岁)过敏性疾病患者,血清检测方法采用欧蒙公司生产的过敏原特异性IgE检测试剂盒。结果：430例患者中,血清过敏原IgE阳性166例(38.60%),其中尘螨和屋尘是主要的吸入性过敏原。血清IgE阳性率男性39%,女性36% (P>0.05),中学组女性阳性率（61.11 %）高于男性（45.83 %）（P<0.05）。不同年龄组间IgE阳性率有统计学差异（P<0.05）,蟑螂、尘螨、霉菌、豚草、屋尘和年龄组间有统计学差异（P<0.05）。不同临床症状与IgE阳性率有统计学差异(P<0.05),其中呼吸道过敏症状组IgE阳性率最高（48.30 %）,不同症状组间主要过敏原都是尘螨。屋尘阳性患者均合并尘螨阳性,其中70.93 %的患者表现出了呼吸道过敏症状。结论：尘螨、屋尘是盐城地区未成年过敏性疾病患者最主要的吸入性过敏原,研究血清过敏原分布,对未成年人过敏性疾病的预防、诊疗具有重要意义。     

Discriminant analysis of lower respiratory tract components associated with cigarette smoking, based on quantitative sputum cytology

Cigarette smoking is the major contributor to chronic bronchial irritation, and tobacco smoke is the most common bronchial carcinogen. A newly developed quantitative sputum cytology test was applied to three-day-pooled, spontaneously produced sputa collected from 349 cigarette smokers and 93 patients who had never smoked; the samples were processed by the Saccomanno technique. In addition to identifying malignant and precancerous cells in sputum, this test identifies eight morphologic parameters of bronchial irritation. Cross-sectional analysis of the cytologic data indicated that, within a 99% confidence interval, cigarette smokers had statistically significantly higher levels of all eight components, producing a unique profile. Discriminant analysis revealed that pigmentation of macrophages, alveolar macrophages and mucous spirals (in that order) were the leading components differentiating smokers from nonsmokers. This new quantitative sputum cytology test correlates with the model of lung carcinogenesis. The detection of proliferative cellular alterations, such as benign and early atypical metaplasia, represent the detection of chronic persistent irritation, which is considered to be a reversible change. The ability to monitor individual responses to toxic inhalants (before the development of irreversible disease) by utilizing a noninvasive test represents a unique opportunity to impact the historic course of lung carcinogenesis.     

Abundance of house dust mites in relation to climate in contrasting agricultural settlements in Israel

The correlation between climatic conditions and mite numbers in houses from rural areas was studied in 13 agricultural communities (kibbutzim and moshavim) in nine geo-climatic subregions of Israel. Mites were present in 97% of the dust samples. The average number of mites per gram of dust in the different localities ranged between 84 and 2053. The maximum number of mites (7440/g dust) was found in a carpet from a house in Geva Carmel in the northern coastal region. The most prevalent species of mites were Dermatophagoides pteronyssinus and Dermatophagoides farinae, which were found in 85.6% and 71.3% of the samples, respectively. The house dust mites D. pteronyssinus, D. farinae and Euroglyphus maynei constituted 94.8% of the mites. Most of the mites were isolated from the carpets and sofas (37.0% and 33.7%, respectively), and a smaller number from beds (29.3%). The smallest number of mites (< or = 250/g dust) were found at a minimum relative humidity (RH) of 30% and lower, with a maximum temperature of 32 degrees C and higher, i.e. in the Jordan valley and Negev mountains. A greater number of mites (250-500/g dust) were found at a minimum ambient RH of 35-40% and a maximum temperature of 32 degrees C and higher, i.e. the Hula valley. A large number of mites (500-1000/g dust) were found at a minimum RH of 35-40% with a maximum temperature of 30 degrees C and lower, i.e. in the Judean and Samarian range, as well as in upper Galilee. The largest number of mites (1000-2000/g dust) was found at a minimum RH of 45% and higher, with a maximum temperature ranging between 30 and 32 degrees C. These conditions occur in the coastal strip, the coastal plain and in the Judean and Samarian foothills. A monthly examination of two houses in Zova, a kibbutz in the Judean hills next to Jerusalem, and two houses from Palmachim, a kibbutz in the coastal region, revealed that the highest prevalence of mites was found in the months April-November and May-November, respectively. In Zova, the highest number of mites were found during the months of June and July while the highest concentrations of D. pteronyssinus-antigen (Der p I) were measured during the month of September. A positive correlation between mite numbers and the quantity of Der p I in house dust was found.     

A human case of house dust mite Tarsonemus floricolus collected from sputum

A 23-year-old medical student showed a positive reaction on a skin test for Paragonimus westermani, and two Tarsonemus floricolus mites were subsequently found by sputum examination and identified morphologically. Our report is the first human case of Tarsonemus floricolus in Korea.     

Occurrence of mites and fungi in the homes of patients with allergic manifestations

This survey was carried out to evaluate mites and moulds concentrations in the homes of patients with allergic manifestations and positive skin-test for mites and/or fungi. The home environments of 277 patients were evaluated by questionnaire, by sampling of airborne fungal spores and by determining the occurrence of mites and moulds in dust samples. Among the 277 patients examined, 83% reacted positively to house dust mites. The fungal allergen most frequently responsible for skin positivity was Alternaria tenuis. The prevalent airborne fungi were Cladosporium, Penicillium and Alternaria, followed by Aspergillus and Aureobasidium. The other genera were found in less than 50% of the homes. The presence of domestic mites in dust was documented in more than 85% of the homes sampled. The occurrence of fungi in dust was generally higher than in the air. The most common genera recovered were Penicillium, Candida, Aspergillus, Aureobasidium, Alternaria and Cladosporium. The homes investigated were divided into two groups, damp and dry, on the basis of the reported presence of wet or damp spots on inside surfaces and moulds growing inside the home. Our results showed that the concentration of mites and moulds in dust was significantly higher (p < 0.01) in damp homes as compared to dry ones.In general, a good agreement between skin positivity and presence of the same allergens in patients' home environment was observed for mites and, among fungi, for Alternaria, Cladosporium and Aspergillus. Our results seem to confirm that the study of mycoaeroflora and the quantification of mites and moulds in dust samples of allergic patients' homes could lead to more precise diagnoses and therefore to better prophylactic and therapeutic programs for each individual patient.     

The influence of household pets on the composition and quantity of allergenic mite fauna within Irish homes: a preliminary investigation

Allergenic mites are responsible for inducing hypersensitive reactions in genetically predisposed people worldwide. Mites in dust from 30 Irish homes with pets (dogs, n = 23; cats, n = 7) were compared with those in 30 homes without pets. House dust mites constituted 78% of all mites recorded, with Dermatophagoides pteronyssinus (Acariformes: Pyroglyphidae) representing 57–72% of mites in furniture and mattresses in both home types compared with only 22% of mites in pet beds. Although storage mites accounted for just 13% of all mites recorded, they represented 46% of mites recorded in pet beds. Median levels of the dust mite allergen Der p 1 (µg/g) in dust samples from mattresses in homes without pets were significantly greater than in mattresses from homes with pets, reflecting the greater densities of D. pteronyssinus found in the former home category. Mite species richness was greater in homes with pets (17 species) than in homes without pets (13 species). This suggests that although the presence of pets can result in a wider variety of epidemiologically important mite species within households, increased competition among mite species may result in a more balanced mite fauna in the home, inhibiting the dominance of any one species and hence lowering allergen‐associated risks.     

Assessment of mite numbers: New methods and results

The determination of mite numbers in dust samples depends on the effectiveness of the sampling procedure. We report on mite isolation from dust samples by flotation distinguishing between living and dead mites. The results are varied and unreproducible; ageing processes are of relevance in the case of dead mites. Living mites can be determined more reliably when using mite mobility. Two methods based on this principle, the mobility test and the heat escape method, are described and discussed. In the case of a natural analogous mite development on carpets, mite numbers ranging from 80 000 to 200 000 mites per m² are found after approx. 3 months. It is shown that the mobility of different mite species varies. The mobility test allows the detection of mites in textile objects, and shows how mites are distributed over these objects. It is demonstrated how the infestation of clothes by mites can be determined.