scholarly journals Magnetic Nanoparticle-Based Biosensing Assay Quantitatively Enhances Acid-Fast Bacilli Count in Paucibacillary Pulmonary Tuberculosis

Biosensors ◽  
2018 ◽  
Vol 8 (4) ◽  
pp. 128 ◽  
Author(s):  
Cristina Gordillo-Marroquín ◽  
Anaximandro Gómez-Velasco ◽  
Héctor Sánchez-Pérez ◽  
Kasey Pryg ◽  
John Shinners ◽  
...  
Keyword(s):  
Pulmonary Tuberculosis ◽  
Magnetic Nanoparticle ◽  
Sputum Sample ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Tb Control ◽  
Control Programs ◽  
Acid Fast Bacilli ◽  
Functionalized Magnetic Nanoparticles ◽  
Colorimetric Biosensing

A new method using a magnetic nanoparticle-based colorimetric biosensing assay (NCBA) was compared with sputum smear microscopy (SSM) for the detection of pulmonary tuberculosis (PTB) in sputum samples. Studies were made to compare the NCBA against SSM using sputum samples collected from PTB patients prior to receiving treatment. Experiments were also conducted to determine the appropriate concentration of glycan-functionalized magnetic nanoparticles (GMNP) used in the NCBA and to evaluate the optimal digestion/decontamination solution to increase the extraction, concentration and detection of acid-fast bacilli (AFB). The optimized NCBA consisted of a 1:1 mixture of 0.4% NaOH and 4% N-acetyl-L-cysteine (NALC) to homogenize the sputum sample. Additionally, 10 mg/mL of GMNP was added to isolate and concentrate the AFB. All TB positive sputum samples were identified with an increased AFB count of 47% compared to SSM, demonstrating GMNP’s ability to extract and concentrate AFB. Results showed that NCBA increased AFB count compared to SSM, improving the grade from “1+” (in SSM) to “2+”. Extending the finding to paucibacillary cases, there is the likelihood of a “scant” grade to become “1+”. The assay uses a simple magnet and only costs $0.10/test. NCBA has great potential application in TB control programs.

scholarly journals Tween 80 Improves the Acid-Fast Bacilli Quantification in the Magnetic Nanoparticle-Based Colorimetric Biosensing Assay (NCBA)

Biosensors ◽  
2022 ◽  
Vol 12 (1) ◽  
pp. 29
Author(s):  
Cristina Gordillo-Marroquín ◽  
Héctor J. Sánchez-Pérez ◽  
Anaximandro Gómez-Velasco ◽  
Miguel Martín ◽  
Karina Guillén-Navarro ◽  
...  
Keyword(s):  
Magnetic Nanoparticle ◽  
Point Of Care ◽  
Diagnostic Technique ◽  
Tween 80 ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Acid Fast Bacilli ◽  
Tb Diagnostics ◽  
Short Time ◽  
Colorimetric Biosensing

Despite its reduced sensitivity, sputum smear microscopy (SSM) remains the main diagnostic test for detecting tuberculosis in many parts of the world. A new diagnostic technique, the magnetic nanoparticle-based colorimetric biosensing assay (NCBA) was optimized by evaluating different concentrations of glycan-functionalized magnetic nanoparticles (GMNP) and Tween 80 to improve the acid-fast bacilli (AFB) count. Comparative analysis was performed on 225 sputum smears: 30 with SSM, 107 with NCBA at different GMNP concentrations, and 88 with NCBA-Tween 80 at various concentrations and incubation times. AFB quantification was performed by adding the total number of AFB in all fields per smear and classified according to standard guidelines (scanty, 1+, 2+ and 3+). Smears by NCBA with low GMNP concentrations (≤1.5 mg/mL) showed higher AFB quantification compared to SSM. Cell enrichment of sputum samples by combining NCBA-GMNP, incubated with Tween 80 (5%) for three minutes, improved capture efficiency and increased AFB detection up to 445% over SSM. NCBA with Tween 80 offers the opportunity to improve TB diagnostics, mainly in paucibacillary cases. As this method provides biosafety with a simple and inexpensive methodology that obtains results in a short time, it might be considered as a point-of-care TB diagnostic method in regions where resources are limited.

Evaluation of the Concentration Sputum Smear Technique for the Laboratory Diagnosis of Pulmonary Tuberculosis

2003 ◽  
Vol 33 (3) ◽  
pp. 160-162 ◽  
Author(s):  
Solomon Gebre-Selassie
Keyword(s):  
Developing Countries ◽  
Pulmonary Tuberculosis ◽  
Laboratory Diagnosis ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Tb Control ◽  
Mycobacterial Diseases ◽  
Control Programmes ◽  
Rate Of Recovery ◽  
Concentration Methods

The microbiological diagnosis of pulmonary tuberculosis (PTB) plays a key role in routine and Tuberculosis (TB) Control Programmes in developing countries. Concentration of acid-fast bacilli (AFB) in clinical specimens is an important step in the laboratory diagnosis of mycobacterial diseases. Microscopy of smears of sputum by direct and after mechanical sedimentation and centrifugation methods followed by treatment with 5% sodium hypochlorite (NaOCl) solution for concentration of the organisms were compared and evaluated. The rate of recovery of AFB from sputum was 8.5%, 25.5% and 38.0% for direct smear microscopy, concentration by sedimentation of NaOCl-treated sputa followed by Ziehl-Neelsen staining and concentration by centrifugation after use of NaOCl respectively. Both the concentration methods by the use of NaOCl solution increased the yield of the AFB by more than threefold compared with the direct microscopy of sputum ( P<0.05). The concentration methods by sedimentation, and centrifugation by the treatment of NaOCl, increased the sensitivity to 75% and 77.9%, respectively, and the specificity to 100% for both techniques. In conclusion, the use of NaOCl in the concentration of AFB in sputum is recommended for use in routine laboratory diagnosis of PTB in developing countries.

scholarly journals A Comparison Of Three Versus Two Sputum Smear Microscopy In A Government Medical College, Patiala, India

1970 ◽  
Vol 8 (2) ◽  
pp. 28-30
Author(s):  
J Kishan ◽  
P Kaur ◽  
A Mahajan ◽  
M Monika ◽  
K Navneet ◽  
...  
Keyword(s):  
Pulmonary Tuberculosis ◽  
Sputum Sample ◽  
Work Load ◽  
Early Morning ◽  
Control Programme ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Medical College ◽  
Two Samples ◽  
National Tuberculosis

Introduction: Under the Revised National Tuberculosis Control Programme of India, three sputum samples are examined and 2 samples positivity criteria are used for labeling the patient as sputum positive pulmonary tuberculosis. Recent studies advocate use of two samples (one spot & one morning) for diagnosis of Tuberculosis. The objective was to compare three versus two sputum smears and to study the relevance of third sputum sample for microscopy in the current practice under Revised National Tuberculosis and Control Programme. Methodology: A study of the laboratory register of the designated microscopic centre for the calendar year 2008 was undertaken. In all 9028 suspects were examined. An analysis of contribution of various sputum samples, S1 (fi rst spot sample), M (early morning) & S2 (second spot) towards diagnosis of Pulmonary Tuberculosis was undertaken. Results: Sputum smear examination results of all the patients examined during 2008 were analyzed. Twelve hundred and eighty eight patients (99.3%) were labeled as smear positive tuberculosis when three sputum samples positivity criteria was considered. By applying two samples and any smear positivity criteria 1296 (99.9%) patients were labeled as sputum smear positive. Among 1296 smears, S1 was positive in 1088 (83.8%) and M in 1293 (99.6%) patients. Early morning sample positivity yield was found higher. Conclusion: Considering 2 samples for examination with at least one morning specimen and one sample positivity criteria, the work load on laboratory can be reduced by 1/3rd without affecting case detection rate. DOI: http://dx.doi.org/10.3126/saarctb.v8i2.5898 SAARCTB 2011; 8(2): 28-30

scholarly journals Improved Diagnosis of Pulmonary Tuberculosis using Bleach (Sodium hypochlorite) Concentration Technique at Tertiary Care Hospital

2021 ◽  
Vol 1 (2) ◽  
pp. 15-22
Author(s):  
Saroj Kumar Thakur ◽  
Vishvesh Prakashchandra Bansal ◽  
Jyotsna Mishra ◽  
M.P. Bansal ◽  
Iswari Sapkota ◽  
...  
Keyword(s):  
Pulmonary Tuberculosis ◽  
Sodium Hypochlorite ◽  
Direct Method ◽  
Tertiary Care ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Care Hospital ◽  
Case Detection ◽  
Acid Fast Bacilli ◽  
Concentration Method

Introduction: In the Revised National Tuberculosis Control Program (RNTCP), microscopic examination of sputum for acid-fast bacilli (AFB) is currently the backbone for the diagnosis of pulmonary tuberculosis. Studies have shown liquefaction and concentration of sputum by 5% sodium hypochlorite is useful in providing increased sensitivity and safety for the handling of specimens. Objective: To assess the utility of the 5% Sodium hypochlorite concentration method in increasing the sensitivity of smear microscopy for detection of AFB for diagnosis of pulmonary tuberculosis. Methods: The study included a total of 1000 sputum samples from 500 patients with suspected pulmonary tuberculosis. Direct smears were prepared from the sputum samples as per RNTCP guidelines. The remaining sputum was used for bleach concentration and smears prepared from the concentrated material. Both smears were stained by Ziehl-Neelsen staining and screened for acid-fast bacilli and graded according to the RNTCP guidelines. Results: A total of 158 samples (15.8%) from 89 patients were positive by a routine direct method whereas by concentration method 236 samples (23.6%) from 143 patients were found positive diagnosing additional 54 patients. The gain in sputum smear positivity of 7.8% over the routine method is highly significant (p=0.0000, χ2= 270) with a 10.8% increase in case detection. Conclusions: Improvement in the sensitivity of smears microscopy will be useful in case detection of tuberculosis especially in resource-poor countries. The increased positivity of microscopy by bleach method indicates that would prove useful if included in the RNTCP to improve case detection. Keywords: Concentration; smear positivity; sodium hypochlorite; tuberculosis.

scholarly journals Concordance of two sputum smear microscopy for diagnosis of presumptive pulmonary tuberculosis in a tertiary care hospital

2017 ◽  
Vol 4 (8) ◽  
pp. 2686
Author(s):  
Star Pala ◽  
Kyrshan G. Lynrah
Keyword(s):  
Pulmonary Tuberculosis ◽  
Tertiary Care Hospital ◽  
Sputum Sample ◽  
Tertiary Care ◽  
Positive Sample ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Care Hospital ◽  
Sputum Smear Microscopy ◽  
Tuberculosis Case

Background: Under revised National tuberculosis control program two sputum samples are to be collected for diagnosis of presumptive pulmonary tuberculosis case. The objective of this study is to find the concordance of both sputum smear microscopy samples at a tertiary care hospital. Methods: Hospital based record was collected from designated microscopy center laboratory register. The data collected were from January 2015 to November 2016. Results: A total of 2117 paired of sputum sample were collected for the year 2015-16 in one of the DMC of a tertiary care hospital. A total of 183 patient (at least one sputum sample) were positive (8.64%) and 1934 were both negative. Among the positive sample concordance for both spot and morning samples were 89% (163/183) and discordance where spot sample positive and morning sample negative was 4.91% (9/183) and where spot sample negative and morning sample positive was 6.01% (11/183). Overall discordance between spot and morning samples were only 0.94% (20/2117). Conclusions: There is a good concordance of two sputum samples. Discordance of two sputum samples were <1% in a tertiary care hospital. One sample may be sufficient for operational purpose for diagnosis of pulmonary Tb. But to have a robust recommendation a RCT will be required to see the extent of discordance by ruling out bias in sputum smear examination.  

scholarly journals Clinical impact and cost analysis of the use of either the Xpert MTB Rif test or sputum smear microscopy in the diagnosis of pulmonary tuberculosis in Rio de Janeiro, Brazil

2018 ◽  
Vol 51 (5) ◽  
pp. 631-637
Author(s):  
Anália Zuleika de Castro ◽  
Adriana Rezende Moreira ◽  
Jaqueline Oliveira ◽  
Paulo Albuquerque Costa ◽  
Carolyne Lalucha Alves Lima Da Graça ◽  
...  
Keyword(s):  
Pulmonary Tuberculosis ◽  
Cost Analysis ◽  
Rio De Janeiro ◽  
Clinical Impact ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Sputum Smear Microscopy

scholarly journals Association between Xpert MTB/RIF cycle threshold values and sputum smear microscopy in patients with pulmonary tuberculosis

2021 ◽  
pp. e20200549
Author(s):  
Gabriela Carpin Pagano1 ◽  
Giovana Rodrigues Pereira1,2 ◽  
Karen Gomes D'Ávila3 ◽  
Luciana Rott Monaiar3 ◽  
Denise Rossato Silva1,3,4
Keyword(s):  
Pulmonary Tuberculosis ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Threshold Values ◽  
Sputum Smear Microscopy ◽  
Cycle Threshold

scholarly journals Optimising fluorescein diacetate sputum smear microscopy for assessing patients with pulmonary tuberculosis

PLoS ONE ◽  
2019 ◽  
Vol 14 (4) ◽  
pp. e0214131 ◽  
Author(s):  
Sumona Datta ◽  
Keren Alvarado ◽  
Robert H. Gilman ◽  
Teresa Valencia ◽  
Christian Aparicio ◽  
...  
Keyword(s):  
Pulmonary Tuberculosis ◽  
Fluorescein Diacetate ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Sputum Smear Microscopy

scholarly journals Sputum Smear Positive Pulmonary Tuberculosis Diagnostic Dropout Rate in Public Health Facilities, Addis Ababa, Ethiopia

2019 ◽  
Vol 2019 ◽  
pp. 1-6
Author(s):  
Daniel Melese Desalegn ◽  
Kumera Terfa Kitila ◽  
Boja Dufera Taddese ◽  
Tinsae Kidanemariam Hailu ◽  
Tariku Takle Dinku ◽  
...  
Keyword(s):  
Public Health ◽  
Pulmonary Tuberculosis ◽  
Addis Ababa ◽  
Dropout Rate ◽  
Sputum Smear ◽  
P Value ◽  
Smear Microscopy ◽  
Sputum Smear Microscopy ◽  
Public Health Facilities ◽  
Smear Positive Pulmonary Tuberculosis

Background. Prolonged laboratory diagnostic process of tuberculosis can lead to failure to complete the diagnosis and increase dropout rate of smear positive pulmonary tuberculosis (PTB) cases. This implies such dropout patients without completing diagnosis are critical as infected individuals remain untreated in the community, providing more opportunities for transmission of the disease and adversely affecting the epidemic. The aim of this research is to determine the level of smear positive PTB diagnosis dropout rate of spot-morning-spot sputum microscopy diagnosis method in public health facilities, in Addis Ababa, Ethiopia. Methods. Retrospective review of patient documents in 13 public health facilities’ TB laboratory in Addis Ababa was conducted from October 2011 to March 2016. Data was computerized using Epi-info software and analysed using SPSS version 20.0 software. Descriptive numerical summaries were used to present the findings. Association between the dropout rate and demographic variables was assessed by Chi-square (X2). Bivariate model using Odds Ratio (OR) with a 95% Confidence Interval (CI) was calculated. P-Value less than 0.05 was taken as statistically significant. Results. Of 41,884 presumptive TB patients registered during the 53 months for laboratory investigation, 5.9% were positive for the first spot sputum smear microscopy. Among these positive cases, 142 (5.8%) and 298 (12.1%) did not come back to the laboratory to submitted early morning and second spot sputum specimens, respectively. The diagnostic dropout for morning sputum specimen in hospitals was 5.6% (58/1039) and in health centres was 5.9% (84/1424). However, higher proportion of dropout for second spot sputum specimen in hospitals was 16.4% (170/1039), compared to the health centres, 8.9% (128/1424). Diagnostic dropout of sputum smear microscopy had no significant association with sociodemographic variable (P value >0.05), while it had significant association with facility type (P value <0.05). Conclusion. In this study smear positive pulmonary tuberculosis diagnostic dropout rate was high compared to WHO reported for the new strategy shift implying the importance of shifting to same-day approach. Hence, shifting from conventional to same day is crucial to minimize the TB diagnostic dropout rate in the study area and other similar settings. Further research is needed/recommended in the local setting to compare the yield and dropout rates between same-day and conventional sputum smear microscopy approach.

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