Evaluation of Auramine O staining and conventional PCR for leprosy diagnosis: A comparative cross-sectional study from Ethiopia

Autor: Kassu Desta, Tsegaye Hailu, Munir H. Idriss, Selfu Girma, Yohannes Tsegaye, Abraham Aseffa, Stewart T. Cole, Philippe Busso, Charlotte Avanzi, Shimelis D. Nigusse, Kidist Bobosha
Jazyk: angličtina
Rok vydání: 2018
Předmět:
Male
Polymerase Chain Reaction
Gastroenterology
030207 dermatology & venereal diseases
chemistry.chemical_compound
0302 clinical medicine
Coloring Agents
slit-skin smear
Eosin
Auramine O
lcsh:Public aspects of medicine
Middle Aged
Infectious Diseases
Real-time polymerase chain reaction
Benzophenoneidum
Ziehl–Neelsen stain
Female
Adult
medicine.medical_specialty
lcsh:Arctic medicine. Tropical medicine
Adolescent
lcsh:RC955-962
030231 tropical medicine
polymerase-chain-reaction
challenges
Haematoxylin
Sensitivity and Specificity
Young Adult
mycobacterium-leprae
03 medical and health sciences
Leprosy
Internal medicine
medicine
Humans
relevance
Aged
Bacteriological Techniques
disease
Staining and Labeling
Diagnostic Tests
Routine
bacilli
business.industry
Public Health
Environmental and Occupational Health
biopsies
lcsh:RA1-1270
Gold standard (test)
DNA extraction
Staining
Cross-Sectional Studies
chemistry
Ethiopia
business
Zdroj: PLoS Neglected Tropical Diseases, Vol 12, Iss 9, p e0006706 (2018)
ISSN: 1935-2735
1935-2727
Popis: Background
Diagnosis of leprosy mainly relies on clinical examination due to the inconsistent sensitivity and poor reproducibility of the current laboratory tests. Utilisation of alternative methods to the standard Ziehl Neelsen (ZN), Fite-Faraco (FF) and Haematoxylin and Eosin (H&E) staining procedures may eventually improve leprosy diagnosis.
Methodology/Principal findings
In this comparative study, the performance of the fluorescent Auramine O (AO) staining and polymerase chain reaction (PCR) was assessed with different skin samples using a combination of ZN, FF and H&E staining as the gold standard. AO, ZN, FF, H&E and PCR tests were performed on slit skin smears (SSS) and/or punch biopsies collected from 141 clinically confirmed leprosy cases and 28 non-leprosy skin samples. DNA was extracted from punch biopsies using two different methods with or without mechanical lysis.
Sensitivities were 87.6%, 59.3% and 77% for H&E, ZN and FF, respectively, whereas it reached 65.5% and 77.9% for AO in SSS and tissue sections and 91.1% for PCR in tissue samples. Morover, samples with low bacillary index, sensitivity of AO staining (61.8%) was similar to FF (60%, p>0.05) and lower than PCR (86.6%, p
Conclusions/Significance
Our results showed that for diagnostic purposes, analysis of skin section is more sensitive than SSS, especially for samples with low bacillary load. AO staining on SSS and tissue sections was not significantly better than other routine diagnostic tests but considerably more user friendly. The sensitivity of PCR was higher than current standard methods and increased when combined with more efficient DNA extraction using mechanical and chemical lysis. Therefore, we recommend AO staining for the diagnosis of leprosy in lower health facilities such as health centres and district hospitals and PCR diagnosis at referral level and research centres.
Databáze: OpenAIRE
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