scholarly journals A comparative study of fluorescent microscopy with Ziehl-Neelsen staining and culture for the diagnosis of pulmonary tuberculosis

1970 ◽  
Vol 7 (3) ◽  
pp. 226-230 ◽  
Author(s):  
S Laifangbam ◽  
HL Singh ◽  
NB Singh ◽  
KM Devi ◽  
NT Singh
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Fluorescence Microscopy ◽  
Pulmonary Tuberculosis ◽  
Fluorescent Microscopy ◽  
False Negative ◽  
Diagnostic Methods ◽  
Case Detection ◽  
Detection Rates ◽  
Auramine O ◽  
The Difference

Background: For developing countries with a large number of cases and financial constraints, evaluation of rapid and inexpensive diagnostic methods has great importance. The bacilli in the sputum can be detected microscopically by ZN stain and fluorochrome stain. Objectives : To study the efficacy of fluorescence microscopy in the diagnosis of pulmonary tuberculosis in comparison to Ziehl-Neelsen staining and culture of sputum samples from patients suspected of pulmonary tuberculosis. Materials and methods : 306 sputum samples collected from 102 patients suspected of pulmonary tuberculosis were processed by the Petroff's method, and subjected to Ziehl-Neelsen staining (ZN), fluorescent Auramine-O staining (AO) and culture on modified Lowenstein-Jensen media (gold standard) for detection of Mycobacterium tuberculosis. Positive smears were graded according to Forbes BA et al, and culture isolates were biochemically tested for confirmation of species. Results : Out of 102 patients, 44.1%, 71.6% and 70% were found positive by ZN, AO and culture respectively. AO was found to be superior to ZN on several aspects. The difference in their case detection rates was statistically significant (χ2 = 24.93, p < 0.001). AO was also able to detect more pauci-bacillary cases than ZN. There was more agreement between culture and fluorescence microscopy (95.1%) than with ZN microscopy (69.6%). The percentage of false negative by AO staining was only 2.78% which was in sharp contrast to that of ZN (40.27%). Conclusion: The better case detection rates of AO over ZN were comparable to those found by several studies. Since screening was done under lower power of magnification (400x), fluorescence microscopy has been found to be less time consuming as compared to ZN method (1000x) in the diagnosis of tuberculosis. The tubercle bacilli stood out as bright objects against a dark background in fluorescence microscopy which makes them easily identifiable hence causing less eye-strain. The efficacy of fluorescence microscopy proved to be much higher than conventional light microscopy and comparable to that of culture. Key words: Ziehl-Neelsen staining; Mycobacterium tuberculosis; Auramine-O DOI: 10.3126/kumj.v7i3.2728 Kathmandu University Medical Journal (2009) Vol.7, No.3 Issue 27, 226-230

scholarly journals Usefulness of Light Emitting Diode (LED) fluorescent microscopy as a tool for rapid and effective method for the diagnosis of pulmonary tuberculosis

1970 ◽  
Vol 37 (1) ◽  
pp. 7-10 ◽  
Author(s):  
Z Khatun ◽  
M Kamal ◽  
CK Roy ◽  
T Sultana ◽  
MQ Rahman ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Pulmonary Tuberculosis ◽  
Light Emitting Diode ◽  
Fluorescent Microscopy ◽  
Infectious Agent ◽  
Sputum Smear ◽  
Light Emitting ◽  
Brightfield Microscopy ◽  
The Difference ◽  
A Prospective Study

Background: Tuberculosis remains world's leading cause of death from a single infectious agent. Fluorescence microscopy offers well-described benefits, comparing with brightfield microscopy, for the evaluation sputum smear samples for tuberculosis. We evaluated the diagnostic performance of fluorescence microscopy, using novel Light Emitting Diode (LED) technology as an alternative to the conventional fluorescence microscopy by Auramine stain as well as brightfield microscopy by Ziehl- Neelsen (ZN) stain.Objectives: The objective of the study was to see the usefulness of LED fluorescent microscopy in the diagnosis of pulmonary tuberculosis.Methods: This is a prospective study consisted of 150 sputum samples from the patients of NIDCH, Mohakhali. All samples were stained by auramine and ZN stain at BSMMU and culture was done in Lowenstein-Jensen (L-J) media as gold standard at NTRL, Mohakhali.Results: In this study total 66(44%) out of 150 sputum specimens were positive for Mycobacterium Tuberculosis by culture. Sensitivity and specificity documented for the different modalities were 95.38% and 94.11%, respectively, for the LED assessment; 68.18% and 90.47%, respectively, for the CFM assessment; and 56.06% and 97.61%, respectively, for brightfield microscopy by ZN stain. The difference in their case detection rate was statistically significant (χ2=119.38, p<0.001).Conclusion: Fluorescence Microscopy (FM) is more sensitive than ZN for diagnosis of pulmonary tuberculosis. However, since FM is more sensitive and rapid, using this method (LED) in clinical laboratories with large specimen numbers is recommended. DOI: http://dx.doi.org/10.3329/bmrcb.v37i1.7792 Bangladesh Med Res Counc Bull 2011; 37: 7 -10

scholarly journals MYCOPLASMAS: BIOLOGY, DISTRIBUTION AND ROLE IN PATHOLOGY

2018 ◽  
Vol 1 (69) ◽  
pp. 50-54
Author(s):  
Виктор Катола ◽  
Viktor Katola
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Mycobacterium Tuberculosis ◽  
Pulmonary Tuberculosis ◽  
Blood Plasma ◽  
Clinical Features ◽  
Diagnostic Methods ◽  
Mycoplasma Infection ◽  
The Family ◽  
Scanning Electron

The general characteristics of the Mollicutes class of the family Mycoplasmataceae, the distribution and sources of mycoplasma infection, its clinical features and diagnostic methods are given. In scanning electron microscopy of blood plasma in patients with severe fibrous cavernous pulmonary tuberculosis in the phase of infiltration and seeding, elementary bodies of unidentified L-forms of bacteria and filamentous branching forms with various structures on the surface, presumably cells of mycoplasmas, were identified. All these formations together with mycobacterium tuberculosis form superinfection, which is the cause of the progression and outcome of the tuberculosis process.

scholarly journals Gen X-Pert MTB/Rif Assay with the Ziehl Nielsen, and Florescence Microscopy Detection Rate of Urinary Tuberculosis Among Suspected and Pulmonary MTB Positive Patients Visiting University of Gondar Specialized Referral Hospital, Northwest Ethiopia.

2021 ◽  
Author(s):  
Dessie Eshetie ◽  
Minichil Worku ◽  
Abiye Tigabu ◽  
Melak Aynalem ◽  
Nega Berhane
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Urinary Tract ◽  
Fluorescence Microscopy ◽  
Pulmonary Tuberculosis ◽  
Detection Rate ◽  
Extra Pulmonary Tuberculosis ◽  
Extra Pulmonary ◽  
Urinary Tuberculosis ◽  
Study Participants ◽  
Gen X

Abstract Background Tuberculosis is a bacterial infection caused by Mycobacterium tuberculosis remains a major global public health concern. Extra-pulmonary tuberculosis accounts for 15% of the global tuberculosis burden. Urinary tract tuberculosis is one of the most common and severe forms of extra-pulmonary tuberculosis in clinical practice. Diagnosis of urinary tract tuberculosis by Gen X-pert MTB/RIF assay from developing countries including Ethiopia is limited. Thus, this study was aimed to compare Gene X-pert MTB/RIF assay with the convectional diagnosis methods. Methods A hospital-based cross-sectional study was conducted among confirmed pulmonary tuberculosis and suspected for urinary tract tuberculosis patients at University of Gondar Specialized Referral Hospital from February 2020 to June 2020 G.C. Non-randomized purposive sampling technique was used to select study participants. To detect Mycobacterium tuberculosis, a urine sample was collected. Then, Ziehl Nielsen and fluorescence microscope, Gene X-pert Real-time PCR were performed to detect the Mycobacterium tuberculosis. Sociodemographic, clinical data, and laboratory data were collected and entered into EPI-Info version 3.5.3 and then transferred to SPSS version-20 for analysis. Descriptive statistics were summarized as percentages, means, and standard deviations. Results A total of 64 study participants were enrolled in this study, 64.2% (41/64) were males and 30% (19/64) were in the age group of 31–45 years. Moreover, 71.9% (46/64) and 57.8% (37/64) study participants were rural residences and illiterate respectively. Among the 64 study participants, 4.69% (3/64) were positive for urinary tuberculosis by Gene X-expert. However, 1.56% (1/64) was positive by fluorescence microscopy, and there was no urinary tuberculosis detected by Ziehl Nielsen examination method. Conclusion and recommendation: The prevalence of urinary tract Mycobacterium tuberculosis using Gene X-pert and fluorescence microscopy was 4.69% (3/64) and 1.56% (1/64), respectively. Gene X-pert has higher detection rate than the conventional methods. Therefore, it is better to develop a guideline on how to use Gen x-pert for the diagnosis for urinary tract tuberculosis in urine samples.

scholarly journals Diagnostic Utility of Bronchial Washing CBNAAT for Mycobacterium Tuberculosis in Sputum Smear Negative and Sputum CBNAAT Negative Patients of Suspected Pulmonary Tuberculosis

2021 ◽  
Vol 6 (03) ◽  
pp. 201-204
Author(s):  
Afshan Ali Shaik ◽  
Uday Kakodkar ◽  
Cigy Borges
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Pulmonary Tuberculosis ◽  
Fluorescent Microscopy ◽  
Case Series ◽  
Tuberculosis Treatment ◽  
Sputum Smear ◽  
Diagnostic Utility ◽  
Fungal Culture ◽  
Bronchial Washing ◽  
Smear Negative

Introduction: Diagnosis of sputum negative pulmonary tuberculosis can be challenging and time consuming, with many patients being put on empirical anti-tuberculosis treatment. Bronchoscopy helps in early diagnosis in such patients. Aims and Objectives: To assess the diagnostic utility of bronchial washing CBNAAT for Mycobacterium Tuberculosis in sputum smear negative and sputum CBNAAT negative patients of suspected pulmonary tuberculosis. Methodology: This case series was conducted on 71 patients in the Department of Pulmonary Medicine, Goa Medical College from May 2018- February 2020. Patients with suspected pulmonary tuberculosis on chest radiograph and/or CT thorax, but with sputum smear as well as CBNAAT for Mycobacterium Tuberculosis negative were subjected to flexible video bronchoscopy. Bronchial washings were collected from the affected lobes and the specimen were subjected to fluorescent microscopy, CBNAAT, MGIT, bacterial and fungal culture. Results: The diagnosis of tuberculosis was established in 23 (32.4%) patients in bronchial washings specimen by CBNAAT. These patients were followed up after 6 months of anti-tuberculosis treatment and have shown significant clinical and radiological improvement. Conclusion: Bronchoscopy is a useful tool in diagnosis of pulmonary tuberculosis in sputum negative patients. It also play an important role in diagnosis of other infectious /malignant disorder which can mimic tuberculosis.

scholarly journals SARS-CoV-2 laboratory diagnostic methods: a systematic review

2021 ◽  
Vol 31 (Supplement_2) ◽  
Author(s):  
Rúben Nunes ◽  
Carolina Melo ◽  
Diana Martins ◽  
Fernando Mendes
Keyword(s):  
Systematic Review ◽  
Point Of Care ◽  
False Negative ◽  
Culture Technique ◽  
Diagnostic Methods ◽  
Blood Collection ◽  
Medical Subject Headings ◽  
Serological Tests ◽  
Detection Rates ◽  
Specimen Handling

Abstract Background The new SARS-CoV-2 is a single-stranded RNA β-coronavirus, and it is comprising by structural proteins (N), (S), (E) and (M). Timely and accurate detection is necessary for spread control of SARS-CoV-2 infection, avoiding inaccurate outcomes. Therefore, in this systematic review, it is purpose to evaluate a set of laboratory diagnostic techniques and methods, analyse their specificity, sensitivity, contributing to improve detection rates and reduce false negative and false positive diagnostic results. Methods In this review, we used literature available on the indexed search database ‘PubMed’, concerning the following keywords: ‘SARS-CoV-2’, ‘Specimen handling’, ‘Cell Culture Technique’, ‘Viral Antigen’, ‘Immunoassay’, ‘NAAT’ according to Medical Subject Headings Mesh and applying the Prisma Diagram methodology. Results DD-PCR and CRISPER had more promising results, but they are unconventional and expensive NAAT methods, the usage of the target RdRp/He gene has exhibited very encouraging results improving sensitivity and specificity. After the onset of symptoms, the sensitivity of the immunoassay varies considering of blood collection day. The antigen detection test showed the highest specificity reached, however is sensitivity is lower compared to NAAT assays. Conclusions In summary, RT-PCR still considered the gold standard and precisely detects the presence of virus RNA, although with some flaws in sensitivity, while the serological tests identify a previous infection by the virus, detecting antibodies, allowing the kinetic and immunological studies, not being able to use as a diagnostic method. The molecular point of care testing equipment’s revelled be the future in the rapid and accurately diagnosis of SARS-CoV-2.

scholarly journals Fluorescence microscopy for the diagnosis of smear-negative pulmonary tuberculosis in Ethiopia

2020 ◽  
Vol 9 (1) ◽  
Author(s):  
Gemeda Abebe ◽  
Dossegnaw Aragaw ◽  
Mulualem Tadesse
Keyword(s):  
Confidence Interval ◽  
Fluorescence Microscopy ◽  
Pulmonary Tuberculosis ◽  
Diagnostic Performance ◽  
Light Emitting Diode ◽  
Fluorescent Microscopy ◽  
Culture Method ◽  
Light Emitting ◽  
Smear Negative ◽  
Negative Sputum

Background: Despite its low sensitivity, microscopy remains the main method for the diagnosis of pulmonary tuberculosis in most laboratories in Ethiopia. Few studies have evaluated the performance of light-emitting diode fluorescent microscopy (LED-FM) in bleach-concentrated smear-negative sputum specimens.Objective: This study aimed to evaluate the diagnostic performance of LED-FM for smear-negative pulmonary tuberculosis in Ethiopia.Methods: A total of 194 adult patients with a cough lasting for more than two weeks, and who had three direct smear-negative sputum tests for Mycobacterium tuberculosis by Ziehl-Neelsen light microscopy, were included. All direct Ziehl-Neelsen-stained smear-negative sputum samples were cultured and were also visualised by LED-FM. Smears for LED-FM were performed from bleach-concentrated sputum sediment. The diagnostic performance of the LED-FM was compared to the culture method (the reference standard).Results: Of the 194 smear-negative sputum specimens analysed, 28 (14.4%) were culture-positive and 21 (10.8%) were LED-FM-positive for M. tuberculosis. However, only 11 of the 21 (52.4%) LED-FM-positive patients had a confirmed tuberculosis diagnosis by culture. Light-emitting diode fluorescence microscopy (FM) had a sensitivity of 39.3% (95% confidence interval: 21.2–57.4) and specificity of 93.9% (95% confidence interval: 90.4–97.6). Ten LED-FM-positive specimens were culture-negative, and all of these specimens had scanty grading (1–19 bacilli per 40 fields on LED-FM).Conclusion: This study showed that implementation of LED-FM on bleach pre-treated and concentrated sputum can significantly improve the diagnosis of smear-negative pulmonary tuberculosis. However, all scanty grade, positive smears by LED-FM need to be confirmed by reference culture method.

scholarly journals Improved detection of Mycobacterium tuberculosis using two independent PCR targets in a tertiary care centre in South India

2011 ◽  
Vol 6 (01) ◽  
pp. 46-52 ◽  
Author(s):  
Ramya Barani ◽  
Gopalsamy Sarangan ◽  
Tessa Antony ◽  
Soundararajan Periyasamy ◽  
Anupma Jyoti Kindo ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
South India ◽  
Tertiary Care ◽  
Fluorescent Microscopy ◽  
Improve Patient Care ◽  
Target Site ◽  
Care Centre ◽  
Tertiary Care Centre ◽  
Auramine O ◽  
Extra Pulmonary

Introduction:  Tuberculosis (TB) causes significant morbidity and mortality worldwide as one of the leading infectious diseases. In India, more than 1.8 million new cases occur every year. Rapid and accurate diagnosis of TB would improve patient care and limit its transmission.This study aimed to evaluate a dual target polymerase chain reaction (PCR) diagnostic assay to detect Mycobacterium tuberculosis from pulmonary and extra-pulmonary samples at a tertiary care centre in South India. Methodology: Samples were collected from patients with a low index of suspicion of TB. Acid-fast smears were performed by Auramine O fluorescent microscopy and PCR was performed by using two site-specific primer pairs targeting IS6110 by nested PCR and TRC4 by conventional PCR. Amplified products for IS6110 and/or TRC4 were indicative of M. tuberculosis.Results: Among 114 (19 pulmonary and 95 extra-pulmonary) samples tested by PCR assay, 12 (11%) were positive for both IS6110 and TRC4, of which 11 (10%) were non-respiratory and one was (1%) respiratory in origin. PCR for TRC4 alone was positive for eight (7%) non-respiratory and two (2%) respiratory samples, while IS6110 alone tested positive for six (5%) non-respiratory samples and one (1%) respiratory sample. Of a total of 29 PCR positive samples, 17 (15 %) were acid-fast smear positive. Conclusion: Although the target site of IS6110 is specific for M. tuberculosis, some strains from South India may lack this region. Therefore, the use of an additional target site (TRC4) is required for improved detection of M. tuberculosis.

scholarly journals Role of cartridge-based nucleic acid amplification test to diagnose tuberculosis at tertiary care teaching hospital in Rajasthan, India

2019 ◽  
Vol 7 (11) ◽  
pp. 4243
Author(s):  
Rajendra Babu Mathur ◽  
Uma Shankar Shukla ◽  
Hemant Kumar Bindal
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Pulmonary Tuberculosis ◽  
Tertiary Care ◽  
Fluorescent Microscopy ◽  
Resistance Rate ◽  
Health Care Facilities ◽  
Rifampicin Resistance ◽  
Smear Negative ◽  
Extra Pulmonary

Background: Tuberculosis is one of the top 10 causes of death worldwide as per the Global TB report 2017, the estimated incidence of TB in India was approximately 28,00,000 cases accounting for about a quarter of the world’s TB cases (10 million). It is of utmost important to diagnose early and treat it to reduce disease transmission. GeneXpert MTB/RIF, an automated cartridge-based molecular technique detects Mycobacterium Tuberculosis and rifampicin resistance within two hours, has been recommended by WHO for rapid diagnosis of TB.Methods: Author conducted a retrospective study in the Department of TB and Chest, of tertiary care center at Jhalawar Medical College (JMC), Jhalawar to evaluate and analyze the role of CBNAAT to diagnose tuberculosis from 1st January 2018 to 31st December 2018. Author included all patients who came to department of TB and Chest of JMC, Jhalawar either new/ relapsed/ defaulters/ referred cases from ART/ ICTC center, Pediatric Department; Gynaecology and Obstetrics Department, peripheral Government Health Care Facilities and Private Hospitals of Jhalawar District catering about 15.5 lac population were subjected to both ZN staining/ Fluorescent microscopy and CBNAAT in the study period.Results: A total of 3078 samples (pulmonary 2739+EP 339) were tested for ZN staining / Fluorescent microscopy and CBNAAT during the study period. Mean age of the study population was 36.5±10.3 years. 1873 tested were negative and 1205 samples were positive for CBNAAT. Of these 1205 positive samples, 1174 were sputum/ BAL samples and 31 were extra pulmonary samples. Authors found rifampicin resistance rate of 6.98% (82/1174) in pulmonary tuberculosis cases, 3 rifampicin resistance cases were detected in extra pulmonary samples. CBNAAT could identify 255 cases (14.01%) that were smear negative. Author found TB-HIV coinfection rate of 18.75%.Conclusions: Author found CBNAAT to be an important diagnostic modality especially in smear negative patients for early diagnosis and treatment. Author could detect Mycobacterium Tuberculosis in 14.01% of patients with negative smear microscopy for AFB. In PLHIV, CBNAAT detected Mycobacterium Tuberculosis in 18.75% (12/64) of patients. Author found rifampicin resistance rate of 6.98% (82/1174) in pulmonary tuberculosis cases.

scholarly journals A Comparision of Laboratory Diagnostic Methods of Tuberculosis and Aetiology of Suspected Cases of Pulmonary Tuberculosis

2015 ◽  
Vol 11 (2) ◽  
pp. 1-6
Author(s):  
Chandra Prakash Bhatt ◽  
B Timalsina ◽  
B Kutu ◽  
R Pradhan ◽  
B Maharjan ◽  
...  
Keyword(s):  
Pulmonary Tuberculosis ◽  
Microscopic Examination ◽  
Fluorescent Microscopy ◽  
Public Health Problem ◽  
Diagnostic Methods ◽  
Fluorescent Staining ◽  
Acid Fast Bacilli ◽  
Direct Microscopic Examination ◽  
Culture Positive ◽  
Culture Negative

Introduction: Tuberculosis remains a worldwide public health problem despite the highly effective drugs and vaccines are available making tuberculosis a preventable and curable disease. The objective of this study was to compare the different laboratory diagnostic methods of tuberculosis and determine its prevalence. Methodology: Morning sputum samples were collected from suspected cases of pulmonary tuberculosis and preceded for Ziehl Neelsen staining, fluorescent staining (auramin-O) and mycobacterium culture in Lowenstein Jensen medium. Results: Total 78 suspected cases of pulmonary tuberculosis were included in this study among them 53 were male and 25 were female. Out of 78 cases, 46 cases were found to be culture positive. In culture positive cases (83%) were found to be M. tuberculosis and (17%) were found to be slow grower, fine colonies, AFB positive but niacin test negative (mycobacteria other than M. tuberculosis). In the direct microscopic examination by Ziehl Neelsen stained smear 26 samples were found to be acid fast bacilli and one sample was culture negative but acid fast bacilli positive. In fluorescent stained smear 34 samples were found to be positive for acid fast bacilli and 5 samples were culture negative but acid fast bacilli positive. Culture was accepted as gold standard, the sensitivity of direct microscopic examination was found 56.5% for Ziehl Neelsen staining and 73.9% for fluorescent staining respectively. Conclusion: In culture positive cases M. tuberculosis and mycobacteria other than M. tuberculosis was found to be 83% and 17% respectively, it was found higher in male than female. Fluorescent microscopy is superior to Ziehl Neelsen microscopy but gives more false positive result than Z-N staining. Combining of Ziehl Neelsen and fluorescent staining is better than fluorescent staining alone.DOI: http://dx.doi.org/10.3126/saarctb.v11i2.12427  SAARC J TUBER LUNG DIS HIV/AIDS, 2014;XI(2), page: 1-6

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