LED fluorescence microscopy for the diagnosis of pulmonary tuberculosis: a multi-country cross-sectional evaluation

Luis Eduardo Cuevas; Najla Al-Sonboli; Lovett Lawson; Mohammed Ahmed Yassin; Isabel Arbide; Nasher Al-Aghbari; Jeevan Bahadur Sherchand; Amin Al-Absi; Emmanuel Nnamdi Emenyonu; Yared Merid; Mosis Ifenyi Okobi; Juliana Olubunmi Onuoha; Melkamsew Aschalew; Abraham Aseffa; Greg Harper; Rachel Mary Anderson de Cuevas; Sally Jane Theobald; Carl-Michael Nathanson; Jean Joly; Brian Faragher; Stephen Bertel Squire; Andrew Ramsay

doi:10.1371/journal.pmed.1001057

LED fluorescence microscopy for the diagnosis of pulmonary tuberculosis: a multi-country cross-sectional evaluation

Luis Eduardo Cuevas, Najla Al-Sonboli, Lovett Lawson, Mohammed Ahmed Yassin, Isabel Arbide, Nasher Al-Aghbari, Jeevan Bahadur Sherchand, Amin Al-Absi, Emmanuel Nnamdi Emenyonu, Yared Merid, Mosis Ifenyi Okobi, Juliana Olubunmi Onuoha, Melkamsew Aschalew, Abraham Aseffa, Greg Harper, Rachel Mary Anderson de Cuevas, Sally Jane Theobald, Carl-Michael Nathanson, Jean Joly, Brian FaragherStephen Bertel Squire, Andrew Ramsay

Research output: Contribution to journal › Article › peer-review

Abstract

BACKGROUND: The diagnosis of tuberculosis (TB) in resource-limited settings relies on Ziehl-Neelsen (ZN) smear microscopy. LED fluorescence microscopy (LED-FM) has many potential advantages over ZN smear microscopy, but requires evaluation in the field. The aim of this study was to assess the sensitivity/specificity of LED-FM for the diagnosis of pulmonary TB and whether its performance varies with the timing of specimen collection.

METHODS AND FINDINGS: Adults with cough ≥2 wk were enrolled consecutively in Ethiopia, Nepal, Nigeria, and Yemen. Sputum specimens were examined by ZN smear microscopy and LED-FM and compared with culture as the reference standard. Specimens were collected using a spot-morning-spot (SMS) or spot-spot-morning (SSM) scheme to explore whether the collection of the first two smears at the health care facility (i.e., "on the spot") the first day of consultation followed by a morning sample the next day (SSM) would identify similar numbers of smear-positive patients as smears collected via the SMS scheme (i.e., one on-the-spot-smear the first day, followed by a morning specimen collected at home and a second on-the-spot sample the second day). In total, 529 (21.6%) culture-positive and 1,826 (74.6%) culture-negative patients were enrolled, of which 1,156 (49%) submitted SSM specimens and 1,199 (51%) submitted SMS specimens. Single LED-FM smears had higher sensitivity but lower specificity than single ZN smears. Using two LED-FM or two ZN smears per patient was 72.8% (385/529, 95% CI 68.8%-76.5%) and 65.8% (348/529, 95% CI 61.6%-69.8%) sensitive (p<0.001) and 90.9% (1,660/1,826, 95% CI 89.5%-92.2%) and 98% (1,790/1,826, 95% CI 97.3%-98.6%) specific (p<0.001). Using three LED-FM or three ZN smears per patient was 77% (408/529, 95% CI 73.3%-80.6%) and 70.5% (373/529, 95% CI 66.4%-74.4%, p<0.001) sensitive and 88.1% (95% CI 86.5%-89.6%) and 96.5% (95% CI 96.8%-98.2%, p<0.001) specific. The sensitivity/specificity of ZN smear microscopy and LED-FM did not vary between SMS and SSM.

CONCLUSIONS: LED-FM had higher sensitivity but, in this study, lower specificity than ZN smear microscopy for diagnosis of pulmonary TB. Performance was independent of the scheme used for collecting specimens. The introduction of LED-FM needs to be accompanied by appropriate training, quality management, and monitoring of performance in the field.

TRIAL REGISTRATION: Current Controlled Trials ISRCTN53339491. Please see later in the article for the Editors' Summary.

Original language	English
Pages (from-to)	e1001057
Journal	PLoS Medicine
Volume	8
Issue number	7
DOIs	https://doi.org/10.1371/journal.pmed.1001057
Publication status	Published - Jul 2011

Keywords

Adult
Clinical Laboratory Techniques
Cough/etiology
Cross-Sectional Studies
Female
Humans
Male
Mass Screening/methods
Microscopy, Fluorescence/methods
Middle Aged
Mycobacterium tuberculosis
Reference Values
Sensitivity and Specificity
Sputum/microbiology
Tuberculosis, Pulmonary/complications
Young Adult

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1371/journal.pmed.1001057

Cite this

Cuevas, L. E., Al-Sonboli, N., Lawson, L., Yassin, M. A., Arbide, I., Al-Aghbari, N., Sherchand, J. B., Al-Absi, A., Emenyonu, E. N., Merid, Y., Okobi, M. I., Onuoha, J. O., Aschalew, M., Aseffa, A., Harper, G., de Cuevas, R. M. A., Theobald, S. J., Nathanson, C.-M., Joly, J., ... Ramsay, A. (2011). LED fluorescence microscopy for the diagnosis of pulmonary tuberculosis: a multi-country cross-sectional evaluation. PLoS Medicine, 8(7), e1001057. https://doi.org/10.1371/journal.pmed.1001057

@article{25dd9916ab564e2eb92e54cb5c31e924,

title = "LED fluorescence microscopy for the diagnosis of pulmonary tuberculosis: a multi-country cross-sectional evaluation",

abstract = "BACKGROUND: The diagnosis of tuberculosis (TB) in resource-limited settings relies on Ziehl-Neelsen (ZN) smear microscopy. LED fluorescence microscopy (LED-FM) has many potential advantages over ZN smear microscopy, but requires evaluation in the field. The aim of this study was to assess the sensitivity/specificity of LED-FM for the diagnosis of pulmonary TB and whether its performance varies with the timing of specimen collection.METHODS AND FINDINGS: Adults with cough ≥2 wk were enrolled consecutively in Ethiopia, Nepal, Nigeria, and Yemen. Sputum specimens were examined by ZN smear microscopy and LED-FM and compared with culture as the reference standard. Specimens were collected using a spot-morning-spot (SMS) or spot-spot-morning (SSM) scheme to explore whether the collection of the first two smears at the health care facility (i.e., {"}on the spot{"}) the first day of consultation followed by a morning sample the next day (SSM) would identify similar numbers of smear-positive patients as smears collected via the SMS scheme (i.e., one on-the-spot-smear the first day, followed by a morning specimen collected at home and a second on-the-spot sample the second day). In total, 529 (21.6%) culture-positive and 1,826 (74.6%) culture-negative patients were enrolled, of which 1,156 (49%) submitted SSM specimens and 1,199 (51%) submitted SMS specimens. Single LED-FM smears had higher sensitivity but lower specificity than single ZN smears. Using two LED-FM or two ZN smears per patient was 72.8% (385/529, 95% CI 68.8%-76.5%) and 65.8% (348/529, 95% CI 61.6%-69.8%) sensitive (p<0.001) and 90.9% (1,660/1,826, 95% CI 89.5%-92.2%) and 98% (1,790/1,826, 95% CI 97.3%-98.6%) specific (p<0.001). Using three LED-FM or three ZN smears per patient was 77% (408/529, 95% CI 73.3%-80.6%) and 70.5% (373/529, 95% CI 66.4%-74.4%, p<0.001) sensitive and 88.1% (95% CI 86.5%-89.6%) and 96.5% (95% CI 96.8%-98.2%, p<0.001) specific. The sensitivity/specificity of ZN smear microscopy and LED-FM did not vary between SMS and SSM.CONCLUSIONS: LED-FM had higher sensitivity but, in this study, lower specificity than ZN smear microscopy for diagnosis of pulmonary TB. Performance was independent of the scheme used for collecting specimens. The introduction of LED-FM needs to be accompanied by appropriate training, quality management, and monitoring of performance in the field.TRIAL REGISTRATION: Current Controlled Trials ISRCTN53339491. Please see later in the article for the Editors' Summary.",

keywords = "Adult, Clinical Laboratory Techniques, Cough/etiology, Cross-Sectional Studies, Female, Humans, Male, Mass Screening/methods, Microscopy, Fluorescence/methods, Middle Aged, Mycobacterium tuberculosis, Reference Values, Sensitivity and Specificity, Sputum/microbiology, Tuberculosis, Pulmonary/complications, Young Adult",

author = "Cuevas, {Luis Eduardo} and Najla Al-Sonboli and Lovett Lawson and Yassin, {Mohammed Ahmed} and Isabel Arbide and Nasher Al-Aghbari and Sherchand, {Jeevan Bahadur} and Amin Al-Absi and Emenyonu, {Emmanuel Nnamdi} and Yared Merid and Okobi, {Mosis Ifenyi} and Onuoha, {Juliana Olubunmi} and Melkamsew Aschalew and Abraham Aseffa and Greg Harper and {de Cuevas}, {Rachel Mary Anderson} and Theobald, {Sally Jane} and Carl-Michael Nathanson and Jean Joly and Brian Faragher and Squire, {Stephen Bertel} and Andrew Ramsay",

year = "2011",

month = jul,

doi = "10.1371/journal.pmed.1001057",

language = "English",

volume = "8",

pages = "e1001057",

journal = "PLoS Medicine",

issn = "1549-1277",

publisher = "Public Library of Science",

number = "7",

}

Cuevas, LE, Al-Sonboli, N, Lawson, L, Yassin, MA, Arbide, I, Al-Aghbari, N, Sherchand, JB, Al-Absi, A, Emenyonu, EN, Merid, Y, Okobi, MI, Onuoha, JO, Aschalew, M, Aseffa, A, Harper, G, de Cuevas, RMA, Theobald, SJ, Nathanson, C-M, Joly, J, Faragher, B, Squire, SB & Ramsay, A 2011, 'LED fluorescence microscopy for the diagnosis of pulmonary tuberculosis: a multi-country cross-sectional evaluation', PLoS Medicine, vol. 8, no. 7, pp. e1001057. https://doi.org/10.1371/journal.pmed.1001057

TY - JOUR

T1 - LED fluorescence microscopy for the diagnosis of pulmonary tuberculosis

T2 - a multi-country cross-sectional evaluation

AU - Cuevas, Luis Eduardo

AU - Al-Sonboli, Najla

AU - Lawson, Lovett

AU - Yassin, Mohammed Ahmed

AU - Arbide, Isabel

AU - Al-Aghbari, Nasher

AU - Sherchand, Jeevan Bahadur

AU - Al-Absi, Amin

AU - Emenyonu, Emmanuel Nnamdi

AU - Merid, Yared

AU - Okobi, Mosis Ifenyi

AU - Onuoha, Juliana Olubunmi

AU - Aschalew, Melkamsew

AU - Aseffa, Abraham

AU - Harper, Greg

AU - de Cuevas, Rachel Mary Anderson

AU - Theobald, Sally Jane

AU - Nathanson, Carl-Michael

AU - Joly, Jean

AU - Faragher, Brian

AU - Squire, Stephen Bertel

AU - Ramsay, Andrew

PY - 2011/7

Y1 - 2011/7

N2 - BACKGROUND: The diagnosis of tuberculosis (TB) in resource-limited settings relies on Ziehl-Neelsen (ZN) smear microscopy. LED fluorescence microscopy (LED-FM) has many potential advantages over ZN smear microscopy, but requires evaluation in the field. The aim of this study was to assess the sensitivity/specificity of LED-FM for the diagnosis of pulmonary TB and whether its performance varies with the timing of specimen collection.METHODS AND FINDINGS: Adults with cough ≥2 wk were enrolled consecutively in Ethiopia, Nepal, Nigeria, and Yemen. Sputum specimens were examined by ZN smear microscopy and LED-FM and compared with culture as the reference standard. Specimens were collected using a spot-morning-spot (SMS) or spot-spot-morning (SSM) scheme to explore whether the collection of the first two smears at the health care facility (i.e., "on the spot") the first day of consultation followed by a morning sample the next day (SSM) would identify similar numbers of smear-positive patients as smears collected via the SMS scheme (i.e., one on-the-spot-smear the first day, followed by a morning specimen collected at home and a second on-the-spot sample the second day). In total, 529 (21.6%) culture-positive and 1,826 (74.6%) culture-negative patients were enrolled, of which 1,156 (49%) submitted SSM specimens and 1,199 (51%) submitted SMS specimens. Single LED-FM smears had higher sensitivity but lower specificity than single ZN smears. Using two LED-FM or two ZN smears per patient was 72.8% (385/529, 95% CI 68.8%-76.5%) and 65.8% (348/529, 95% CI 61.6%-69.8%) sensitive (p<0.001) and 90.9% (1,660/1,826, 95% CI 89.5%-92.2%) and 98% (1,790/1,826, 95% CI 97.3%-98.6%) specific (p<0.001). Using three LED-FM or three ZN smears per patient was 77% (408/529, 95% CI 73.3%-80.6%) and 70.5% (373/529, 95% CI 66.4%-74.4%, p<0.001) sensitive and 88.1% (95% CI 86.5%-89.6%) and 96.5% (95% CI 96.8%-98.2%, p<0.001) specific. The sensitivity/specificity of ZN smear microscopy and LED-FM did not vary between SMS and SSM.CONCLUSIONS: LED-FM had higher sensitivity but, in this study, lower specificity than ZN smear microscopy for diagnosis of pulmonary TB. Performance was independent of the scheme used for collecting specimens. The introduction of LED-FM needs to be accompanied by appropriate training, quality management, and monitoring of performance in the field.TRIAL REGISTRATION: Current Controlled Trials ISRCTN53339491. Please see later in the article for the Editors' Summary.

AB - BACKGROUND: The diagnosis of tuberculosis (TB) in resource-limited settings relies on Ziehl-Neelsen (ZN) smear microscopy. LED fluorescence microscopy (LED-FM) has many potential advantages over ZN smear microscopy, but requires evaluation in the field. The aim of this study was to assess the sensitivity/specificity of LED-FM for the diagnosis of pulmonary TB and whether its performance varies with the timing of specimen collection.METHODS AND FINDINGS: Adults with cough ≥2 wk were enrolled consecutively in Ethiopia, Nepal, Nigeria, and Yemen. Sputum specimens were examined by ZN smear microscopy and LED-FM and compared with culture as the reference standard. Specimens were collected using a spot-morning-spot (SMS) or spot-spot-morning (SSM) scheme to explore whether the collection of the first two smears at the health care facility (i.e., "on the spot") the first day of consultation followed by a morning sample the next day (SSM) would identify similar numbers of smear-positive patients as smears collected via the SMS scheme (i.e., one on-the-spot-smear the first day, followed by a morning specimen collected at home and a second on-the-spot sample the second day). In total, 529 (21.6%) culture-positive and 1,826 (74.6%) culture-negative patients were enrolled, of which 1,156 (49%) submitted SSM specimens and 1,199 (51%) submitted SMS specimens. Single LED-FM smears had higher sensitivity but lower specificity than single ZN smears. Using two LED-FM or two ZN smears per patient was 72.8% (385/529, 95% CI 68.8%-76.5%) and 65.8% (348/529, 95% CI 61.6%-69.8%) sensitive (p<0.001) and 90.9% (1,660/1,826, 95% CI 89.5%-92.2%) and 98% (1,790/1,826, 95% CI 97.3%-98.6%) specific (p<0.001). Using three LED-FM or three ZN smears per patient was 77% (408/529, 95% CI 73.3%-80.6%) and 70.5% (373/529, 95% CI 66.4%-74.4%, p<0.001) sensitive and 88.1% (95% CI 86.5%-89.6%) and 96.5% (95% CI 96.8%-98.2%, p<0.001) specific. The sensitivity/specificity of ZN smear microscopy and LED-FM did not vary between SMS and SSM.CONCLUSIONS: LED-FM had higher sensitivity but, in this study, lower specificity than ZN smear microscopy for diagnosis of pulmonary TB. Performance was independent of the scheme used for collecting specimens. The introduction of LED-FM needs to be accompanied by appropriate training, quality management, and monitoring of performance in the field.TRIAL REGISTRATION: Current Controlled Trials ISRCTN53339491. Please see later in the article for the Editors' Summary.

KW - Adult

KW - Clinical Laboratory Techniques

KW - Cough/etiology

KW - Cross-Sectional Studies

KW - Female

KW - Humans

KW - Male

KW - Mass Screening/methods

KW - Microscopy, Fluorescence/methods

KW - Middle Aged

KW - Mycobacterium tuberculosis

KW - Reference Values

KW - Sensitivity and Specificity

KW - Sputum/microbiology

KW - Tuberculosis, Pulmonary/complications

KW - Young Adult

U2 - 10.1371/journal.pmed.1001057

DO - 10.1371/journal.pmed.1001057

M3 - Article

C2 - 21765809

SN - 1549-1277

VL - 8

SP - e1001057

JO - PLoS Medicine

JF - PLoS Medicine

IS - 7

ER -

LED fluorescence microscopy for the diagnosis of pulmonary tuberculosis: a multi-country cross-sectional evaluation

Abstract

Keywords

UN SDGs

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