Detection of Mycobacterium tuberculosis Complex in Formalin-Fixed, Paraffin-Embedded Tissue Specimens with Necrotizing Granulomatous Inflammation by Strand Displacement Amplification

Johansen, Işik Somuncu; Thomsen, Vibeke Østergaard; Forsgren, Arne; Hansen, Birgit Fischer; Lundgren, Bettina

doi:10.1016/s1525-1578(10)60515-6

Cited by 30 publications

(26 citation statements)

References 29 publications

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“…Nevertheless, the results differ from those obtained by JOHANSEN et al 16 , who described good results with the use of 10% non-buffered formalin. It is possible that the quality of the water used for formalin dilution or other unknown factors might have contributed to these discrepant results.…”

Section: Discussioncontrasting

confidence: 94%

“…Similar results were obtained by GREER et al 7 , who evaluated different fixation times (1h, 4h and 24h) and by JOHANSEN et al 16 who obtained excellent results using fixation times between 16h and 18h. In our experiment, the samples were collected and fixed in less than 24h, which may have contributed to the high positivity in the human betaactin gene amplification.…”

Section: Discussionsupporting

confidence: 85%

“…False negative results of IS6110 PCR might have resulted from a low efficiency of the mycobacterial cell wall lysis or uneven distribution of bacilli in the samples 1,16 . In addition, for the testing, only part of biopsy samples were analysed.…”

Section: Discussionmentioning

confidence: 99%

“…Nevertheless, there are difficulties in obtaining good results using PCR in FFPE tissue 16 . PCR efficiency in FFPE samples may be influenced by several factors, which affect tissue integrity 7,22 , such as the fixation protocol, the age of the paraffin block, and the presence of endogenous or exogenous inhibitors of the reaction 1,7 .…”

mentioning

confidence: 99%

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Effects of tissue handling and processing steps on PCR for detection of Mycobacterium tuberculosis in formalin-fixed paraffin-embedded samples

Barcelos

Franco

Leão

2008

Rev. Inst. Med. trop. S. Paulo

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sUMMarYDevelopment and standardization of reliable methods for detection of Mycobacterium tuberculosis in clinical samples is an important goal in laboratories throughout the world. In this work, lung and spleen fragments from a patient who died with the diagnosis of miliary tuberculosis were used to evaluate the influence of the type of fixative as well as the fixation and paraffin inclusion protocols on PCR performance in paraffin embedded specimens. Tissue fragments were fixed for four h to 48 h, using either 10% non-buffered or 10% buffered formalin, and embedded in pure paraffin or paraffin mixed with bee wax. Specimens were submitted to PCR for amplification of the human beta-actin gene and separately for amplification of the insertion sequence IS6110, specific from the M. tuberculosis complex. Amplification of the beta-actin gene was positive in all samples. No amplicons were generated by PCR-IS6110 when lung tissue fragments were fixed using 10% non-buffered formalin and were embedded in paraffin containing bee wax. In conclusion, combined inhibitory factors interfere in the detection of M. tuberculosis in stored material. It is important to control these inhibitory factors in order to implement molecular diagnosis in pathology laboratories.

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Section: Discussioncontrasting

confidence: 94%

Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Effects of tissue handling and processing steps on PCR for detection of Mycobacterium tuberculosis in formalin-fixed paraffin-embedded samples

Barcelos

Franco

Leão

2008

Rev. Inst. Med. trop. S. Paulo

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“…debe ser rápido y confiable para iniciar un tratamiento oportuno, evitar complicaciones graves y prevenir la transmisión de la enfermedad 1 . Las lesiones granulomatosas características de TBC también ocurren en otras enfermedades infecciosas (brucelosis, lepra, enfermedad del rasguño de gato, en algunos hongos y virus) 2 y no infecciosas (carcinomas, linfomas, sarcoidosis, cuadros tóxicos, alérgicos y autoinmunes) [3][4][5] .…”

Section: E L Diagnóstico De Tuberculosis (Tbc) Causada Por El Complejunclassified

Eficacia del diagnóstico diferencial de tuberculosis mediante reacción de polimerasa en cadena en lesiones granulomatosas de tejidos fijados en formalina e incluidos en parafina

Montenegro¹,

Delgado

Pineda³

et al. 2014

Rev. chil. infectol.

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Development of a Highly Specific, Selective, and Sensitive Fluorescent Probe for Detection of Mycobacteria in Human Tissues

Gupta

Mishra

Khan

et al. 2022

Adv Healthcare Materials

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Tuberculosis (TB), including extrapulmonary TB, is responsible for more than one million deaths in a year worldwide. Existing methods of mycobacteria detection have poor sensitivity, selectivity, and specificity, especially in human tissues. Herein, the synthesis of a cholic acid‐derived fluorescent probe (P4) that can specifically stain the mycobacterium species is presented. It is shown that P4 probe specifically binds with mycobacterial lipids, trehalose monomycolate, and phosphatidylinositol mannoside 6. P4 probe can detect mycobacteria in polymicrobial planktonic cultures and biofilms with high specificity, selectivity, and sensitivity. Moreover, it can detect a single mycobacterium in the presence of 10 000 other bacilli. Unlike the probes that depend on active mycobacterial enzymes, the membrane‐specific P4 probe can detect mycobacteria even in formalin‐fixed paraffin‐embedded mice and human tissue sections. Therefore, the ability of the P4 probe to detect mycobacteria in different biological milieu makes it a potential candidate for diagnostic and prognostic applications in clinical settings.

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Detection of Mycobacterium tuberculosis Complex in Formalin-Fixed, Paraffin-Embedded Tissue Specimens with Necrotizing Granulomatous Inflammation by Strand Displacement Amplification

Cited by 30 publications

References 29 publications

Effects of tissue handling and processing steps on PCR for detection of Mycobacterium tuberculosis in formalin-fixed paraffin-embedded samples

Effects of tissue handling and processing steps on PCR for detection of Mycobacterium tuberculosis in formalin-fixed paraffin-embedded samples

Eficacia del diagnóstico diferencial de tuberculosis mediante reacción de polimerasa en cadena en lesiones granulomatosas de tejidos fijados en formalina e incluidos en parafina

Development of a Highly Specific, Selective, and Sensitive Fluorescent Probe for Detection of Mycobacteria in Human Tissues

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