A Reevaluation of Sputum Microscopy and Culture in the Diagnosis of Pulmonary Tuberculosis

Levy, Howard; Feldman, Charles; Sacho, H; Meulen, Hermina van der; Kallenbach, J; Koornhof, H. J.

doi:10.1378/chest.95.6.1193

Cited by 113 publications

(57 citation statements)

References 7 publications

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“…In smear-positive cases, this approach required less than 2 weeks instead of 8-12 weeks with the standard solid media methods [8,23,24]. Furthermore, in smear-negative cases, this approach reduced the time needed for drug testing using different types of specimens by nearly half, from more than 10 weeks to 5 weeks when solid media were used for primary culture.…”

Section: Discussionmentioning

confidence: 99%

Rapid drug susceptibility testing of Mycobacterium tuberculosis using conventional solid media

Schaberg

Reichert²,

Schülin³

et al. 1995

Eur Respir J

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R Ra ap pi id d d dr ru ug g s su us sc ce ep pt ti ib bi il li it ty y t te es st ti in ng g o of f M My yc co ob ba ac ct te er ri iu um mt tu ub be er rc cu ul lo os si is s u us si in ng g c co on nv ve en nt ti io on na al l s so ol li id d m me ed di ia a In 64 smear-positive patients, the median time for final susceptibility results was 11 days (95% confidence interval (95% CI) 10-12 days) compared to 62 days (95% CI 56-66 days) with the standard method. In 133 smear-negative patients, results were available after a median of 35 days (95% CI 32-40 days) in contrast to 72 days (95% CI 62-83 days) with the standard method, regardless of whether or not sputum or other materials were used for primary culture. The rapid method detected all cases of single-drug resistance (n=20) and multidrug resistance (n=14) within 13 days (95% CI 9-17 days) in smear-positive patients (n=8), or within 38 days (95% CI 35-48 days) in smear-negative patients (n=26). Only one discrepancy was encountered in 985 resistance tests. Moreover, contamination was not observed.Our rapid susceptibility testing method for M. tuberculosis on Middlebrook 7H11 agar is fast, practical and inexpensive. It provides an alternative when more sophisticated techniques are not available or affordable.

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Section: Discussionmentioning

confidence: 99%

Rapid drug susceptibility testing of Mycobacterium tuberculosis using conventional solid media

Schaberg

Reichert²,

Schülin³

et al. 1995

Eur Respir J

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“…In the presence of 10-100 bacilli in a milliliter of the sample and culture, a positive result can be obtained. The sensitivity of sputum culture is around 81.5% and the specificity is around 98.4% (14). Solid mediums such as Löwenstein-Jensen or liquid mediums such as Middlebrook H12 are used for culture.…”

Section: A Microbiologymentioning

confidence: 99%

Smear-Negative Pulmonary Tuberculosis

Çalışkan¹,

Kaya²

2015

Eurasian J Pulmonol

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Tuberculosis (TB) is the second leading cause of death from an infectious disease worldwide, after the human immunodeficiency virus. There are almost 2.5 million new smear-positive pulmonary TB cases and 1.9 million new smear-negative pulmonary TB cases in the world in 2012 according to the World Health Organization (WHO) Global Tuberculosis Report 2013. Smear negativity in pulmonary TB is a common clinical problem. Clinicians have difficulty in diagnosing smear-negative pulmonary TB without bacteriological confirmation. It is very important to decide whether or not to treat a patient with smear negative pulmonary TB when the culture results are pending or negative. New diagnostic methods are required for the diagnosis of smear-negative pulmonary TB. In addition to the development of new microbiological and serological diagnostic tests, clinical prediction scoring systems and algorithms including clinical and radiological findings of smear-negative pulmonary TB patients in our country, should be established to facilitate the diagnosis of smear-negative pulmonary TB.

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“…11,[14][15][16][17] Although this study did not evaluate the influence of concurrent HIV infection on the diagnostic capabilities of LED fluorescence microscopy, we presume that the sensitivity in a person with a co-infection would be improved because smears are often reported as negative because of a low bacillary content. 11,16,[18][19][20] Mycobacterial culture with Löwenstein-Jensen medium is not used for routine diagnosis of TB but it is still considered a gold standard even though this method is time-consuming. 10,18,21 Culture with Löwenstein-Jensen medium was only used to determine discrepancies.…”

mentioning

confidence: 99%

The CyScope® Fluorescence Microscope, a Reliable Tool for Tuberculosis Diagnosis in Resource-Limited Settings

Lehman¹,

Yamadji²,

Sack³

et al. 2010

The American Society of Tropical Medicine and Hygiene

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In the context of human immunodeficiency virus/acquired immunodeficiency syndrome pandemics, tuberculosis (TB) is the most common opportunistic infection. More than 9 million cases in 2007 were found in Africa and Southeast Asia. The absence of accurate diagnostic techniques constitutes a serious hindrance in the strategy of TB control in Africa. 2 The increase of false negative-smear pulmonary TB is highlighted as one of the main reasons for the upsurge of this disease during the past decade in poor countries. 3 In an effort to improve methods to diagnose pulmonary tuberculosis, the World Health Organization has encouraged the development of simplified and accurate diagnostics, such as fluorescence microscopy for early detection of Mycobacterium tuberculosis in sputum. 4 The CyScope ® (Partec, Görlitz, Germany) belongs to a new generation of fluorescence microscopes that use light-emitting diodes (LEDs) as a light source. 5 It can be plugged into an ordinary electrical outlet or operated with a built-in rechargeable battery. It is equipped with a high power Royal Blue LED (wavelength = 455 nm) for incidence fluorescence excitation and a white light LED for transmitted light. The LED fluorescence microscope provided similar results as standard mercury vapor lamp fluorescence microscope at a much lower cost. 6 In this study, we used the LED fluorescence microscope CyScope ® for TB diagnosis and investigated different staining procedures to shorten the handling time for samples.During August-November 2009, 300 sputum samples were collected from patients in Cameroon with suspected pulmonary TB or patients receiving treatment. The age range of the patients was 2-74 years. Two slides were prepared from the same specimen for direct sputum smears by using the unconcentrated specimens in the solid or most dense particles of the sputum. 7 The smears were dried in air and heat-fixed. Staining was classically processed as described by the World Health Organization and the International Union Against Tuberculosis and Lung Disease. A solution with 0.3% basic fuchsin (pararosaniline chloride with 88% P1528 dye; Sigma, St. Louis, MO) was heated to the steaming point, decolorized with 25% sulfuric acid, and counterstained with 0.3% methylene blue. 7,8 The fluorescent method of Degommier 9 was applied. The staining reagent Tb-fluor (Merck, Darmstadt, Germany) was used according to the manufacturer's procedure. Briefly, smears were stained with auramine-rhodamine, rinsed with tap water, decolorized with HCl-isopropanol, and counterstained with KMnO 4 . Five protocols with variable staining and counterstaining durations were used to establish overall minimal staining time with the fluorescent dye. ( Table 1 ).Acid-fast-stained smears were colored by a variety of protocols and observed on the CyScope ® . This microscope uses white light and fluorescence. When the fluorescence function was turned on, the acid-fast bacilli were visualized at magnifications of 200× and 400×. Their number in the expectoration fluid was related to the s...

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A Reevaluation of Sputum Microscopy and Culture in the Diagnosis of Pulmonary Tuberculosis

Cited by 113 publications

References 7 publications

Rapid drug susceptibility testing of Mycobacterium tuberculosis using conventional solid media

Rapid drug susceptibility testing of Mycobacterium tuberculosis using conventional solid media

Smear-Negative Pulmonary Tuberculosis

The CyScope® Fluorescence Microscope, a Reliable Tool for Tuberculosis Diagnosis in Resource-Limited Settings

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