Fluorochrome Staining of Mycobacteria

Weiser, O. L.; Sproat, Erma F.; Hakes, James D.; Morse, Warren C.

doi:10.1093/ajcp/46.5_ts.587

Cited by 4 publications

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“…The decontaminated feces specimen and tissue biopsy were used for three different investigations. First, the detection of AFB was performed via auramine acid-fast staining under fluorescence microscopy [ 17 ]. Second, the specimens were subjected to direct extraction of total DNA via magnetic bead-based method using MagDEA ® Dx SV reagent (Precision System Science, Chiba, Japan) according to the manufacturer’s instructions.…”

Section: Methodsmentioning

confidence: 99%

Clinical characteristics and diagnosis of intestinal tuberculosis in clinical practice at Thailand’s largest national tertiary referral center: An 11-year retrospective review

et al. 2023

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Background Diagnosing intestinal tuberculosis (ITB) is challenging due to the low diagnostic sensitivity of current methods. This study aimed to assess the clinical characteristics and diagnosis of ITB at our tertiary referral center, and to explore improved methods of ITB diagnosis. Methods This retrospective study included 177 patients diagnosed with ITB at Siriraj Hospital (Bangkok, Thailand) during 2009–2020. Results The mean age was 49 years, 55.4% were male, and 42.9% were immunocompromised. Most diagnoses (108/177) were made via colonoscopy; 12 patients required more than one colonoscopy. Among those, the sensitivity of tissue acid-fast bacilli (AFB), presence of caseous necrosis, polymerase chain reaction (PCR), and culture was 40.7%, 13.9%, 25.7%, and 53.4%, respectively. Among patients with negative tissue histopathology, 4 (3.7%) and 13 (12.0%) were ITB positive on tissue PCR and culture, respectively. The overall sensitivity when all diagnostic methods were used was 63%. Seventy-six patients had stool tests for mycobacteria. The overall sensitivity of stool tests was 75.0%. However, when analyzing the 31 patients who underwent both endoscopy and stool testing, the sensitivity of stool testing when using tissue biopsy as a reference was 45.8%. Combining stool testing and tissue biopsy did not significantly increase the sensitivity compared to tissue biopsy alone (83.9% vs. 77.4%, respectively). Conclusion Despite the availability of PCR and culture for TB, the overall diagnostic sensitivity was found to be low. The sensitivity increased when the tests were used in combination. Repeated colonoscopy may be beneficial. Adding stool mycobacteria tests did not significantly increase the diagnostic yield if endoscopy was performed, but it could be beneficial if endoscopy is unfeasible.

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Section: Methodsmentioning

confidence: 99%

Clinical characteristics and diagnosis of intestinal tuberculosis in clinical practice at Thailand’s largest national tertiary referral center: An 11-year retrospective review

et al. 2023

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“…This is particularly true in children (Getahun et al, 2007 ) and HIV-co-infected patients (Swai et al, 2011 ). Alternatively, M. tuberculosis can be stained in bright yellow-orange by fluorescent auramine O or auramine–rhodamine (Figure 1 ; Weiser et al, 1966 ). Observations are made at X 250 using fluorescent light-emitting diode microscopy (LED-FM) which can be operated on batteries for field use (Anthony et al, 2006 ; World Health Organization, 2011 ; Marzouk et al, 2013 ).…”

Section: Introductionmentioning

confidence: 99%

Rapid culture-based diagnosis of pulmonary tuberculosis in developed and developing countries

Asmar

Drancourt

2015

Front. Microbiol.

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Culturing Mycobacterium tuberculosis remains the gold standard for the laboratory diagnosis of pulmonary tuberculosis, with 9 million new cases and 1.5 million deaths mainly in developing countries. Reviewing data reported over 20 years yields a state-of-the-art procedure for the routine culture of M. tuberculosis in both developed and developing countries. Useful specimens include sputum, induced sputum, and stools collected in quaternary ammonium preservative-containing sterile cans. The usefulness of other non-invasive specimens remains to be evaluated. Specimens can be collected in a diagnosis kit also containing sampling materials, instructions, laboratory requests, and informed consent. Automated direct LED fluorescence microscopy after auramine staining precedes inoculation of an egg-lecithin-containing culture solid medium under microaerophilic atmosphere, inverted microscope reading or scanning video-imaging detection of colonies and colonies identification by recent molecular methods. This procedure should result in a diagnosis of pulmonary tuberculosis as fast as 5 days. It may be implemented in both developed and developing countries with automated steps replaceable by manual steps depending on local resources.

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Acridine orange as a fluorescent counterstain with the auramine acid-fast stain

Smithwick

David

1971

Tubercle

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Fluorochrome Staining of Mycobacteria

Cited by 4 publications

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Clinical characteristics and diagnosis of intestinal tuberculosis in clinical practice at Thailand’s largest national tertiary referral center: An 11-year retrospective review

Clinical characteristics and diagnosis of intestinal tuberculosis in clinical practice at Thailand’s largest national tertiary referral center: An 11-year retrospective review

Rapid culture-based diagnosis of pulmonary tuberculosis in developed and developing countries

Acridine orange as a fluorescent counterstain with the auramine acid-fast stain

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