Mycobacterium tuberculosis complex detected by modified fluorescent in situ hybridization in lymph nodes of clinical samples
Introduction: Lymph node tuberculosis (TB) is the leading cause of extrapulmonary tuberculosis and is the most frequently identified type in Aguascalientes, Mexico. Conventional diagnosis has serious limitations for rapid detection of extrapulmonary tuberculosis in clinical samples. Here PCR and mo...
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The Journal of Infection in Developing Countries
2011-11-01
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| Series: | Journal of Infection in Developing Countries |
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| Online Access: | https://jidc.org/index.php/journal/article/view/1752 |
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| author | Juan Rodriguez-Nuñez Francisco J Avelar Francisco Marquez Bruno Rivas-Santiago Cesar Quiñones Alma L. Guerrero-Barrera |
| author_facet | Juan Rodriguez-Nuñez Francisco J Avelar Francisco Marquez Bruno Rivas-Santiago Cesar Quiñones Alma L. Guerrero-Barrera |
| author_sort | Juan Rodriguez-Nuñez |
| collection | DOAJ |
| description | Introduction: Lymph node tuberculosis (TB) is the leading cause of extrapulmonary tuberculosis and is the most frequently identified type in Aguascalientes, Mexico. Conventional diagnosis has serious limitations for rapid detection of extrapulmonary tuberculosis in clinical samples. Here PCR and modified FISH have been tested as complementary diagnosis methods for extrapulmonary tuberculosis.
Methodology: The specific insertion sequence IS6110 for Mycobacterium tuberculosis complex was used to perform PCR and build DNA and PNA FISH probes (20bp). PCR and modified DNA and PNA FISH assays were performed to evaluate 41 lymph node paraffin-embedded tissue samples, in comparison with the histopathology diagnosis, which was considered the gold standard (22 positive and 19 negative).
Results: In comparison with histopathology diagnosis PCR showed 62.5 % sensitivity and 77.8 % specificity (χ2 = 4.583 p < 0.05). Modified DNA FISH showed 71.4% sensitivity and 84.6% specificity (χ2 = 11.21 p < 0.05). PNA FISH showed 66.7% sensitivity and 60.0% specificity (χ2 = 2.93 p > 0.05). Ziehl Neelsen stain was positive in only four cases of 22 lymph node samples positive to histopathology. In contrast, PCR and modified DNA FISH were positive in 20 cases of the same group. The negative cases were coincident in all tests.
Conclusions: PCR and DNA FISH showed a significant increase in the number of cases detected and also showed higher sensitivity and specificity compared with data reported by traditional methodology. In developing countries, these techniques could help to complement the early diagnosis and timely treatment of extrapulmonary tuberculosis.
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| format | Article |
| id | doaj-art-e1c96d0bccb845a7ba00078056413c43 |
| institution | OA Journals |
| issn | 1972-2680 |
| language | English |
| publishDate | 2011-11-01 |
| publisher | The Journal of Infection in Developing Countries |
| record_format | Article |
| series | Journal of Infection in Developing Countries |
| spelling | doaj-art-e1c96d0bccb845a7ba00078056413c432025-08-20T02:16:15ZengThe Journal of Infection in Developing CountriesJournal of Infection in Developing Countries1972-26802011-11-0160110.3855/jidc.1752Mycobacterium tuberculosis complex detected by modified fluorescent in situ hybridization in lymph nodes of clinical samplesJuan Rodriguez-Nuñez0Francisco J Avelar1Francisco Marquez2Bruno Rivas-Santiago3Cesar Quiñones4Alma L. Guerrero-Barrera5Universidad Autónoma de Aguascalientes, Aguascalientes, MéxicoUniversidad Autónoma de Aguascalientes, Aguascalientes, MéxicoCentenario Hospital Miguel Hidalgo, Aguascalientes, MéxicoUnidad de Investigación Médica-Zacatecas México, IMSSUniversidad Autónoma de Aguascalientes, Aguascalientes, MéxicoUniversidad Autónoma de Aguascalientes, Aguascalientes, MéxicoIntroduction: Lymph node tuberculosis (TB) is the leading cause of extrapulmonary tuberculosis and is the most frequently identified type in Aguascalientes, Mexico. Conventional diagnosis has serious limitations for rapid detection of extrapulmonary tuberculosis in clinical samples. Here PCR and modified FISH have been tested as complementary diagnosis methods for extrapulmonary tuberculosis. Methodology: The specific insertion sequence IS6110 for Mycobacterium tuberculosis complex was used to perform PCR and build DNA and PNA FISH probes (20bp). PCR and modified DNA and PNA FISH assays were performed to evaluate 41 lymph node paraffin-embedded tissue samples, in comparison with the histopathology diagnosis, which was considered the gold standard (22 positive and 19 negative). Results: In comparison with histopathology diagnosis PCR showed 62.5 % sensitivity and 77.8 % specificity (χ2 = 4.583 p < 0.05). Modified DNA FISH showed 71.4% sensitivity and 84.6% specificity (χ2 = 11.21 p < 0.05). PNA FISH showed 66.7% sensitivity and 60.0% specificity (χ2 = 2.93 p > 0.05). Ziehl Neelsen stain was positive in only four cases of 22 lymph node samples positive to histopathology. In contrast, PCR and modified DNA FISH were positive in 20 cases of the same group. The negative cases were coincident in all tests. Conclusions: PCR and DNA FISH showed a significant increase in the number of cases detected and also showed higher sensitivity and specificity compared with data reported by traditional methodology. In developing countries, these techniques could help to complement the early diagnosis and timely treatment of extrapulmonary tuberculosis. https://jidc.org/index.php/journal/article/view/1752fluorescent in-situ hybridizationFISHtuberculosisMycobacterium tuberculosis complexPCRinsertion sequence 6110 |
| spellingShingle | Juan Rodriguez-Nuñez Francisco J Avelar Francisco Marquez Bruno Rivas-Santiago Cesar Quiñones Alma L. Guerrero-Barrera Mycobacterium tuberculosis complex detected by modified fluorescent in situ hybridization in lymph nodes of clinical samples Journal of Infection in Developing Countries fluorescent in-situ hybridization FISH tuberculosis Mycobacterium tuberculosis complex PCR insertion sequence 6110 |
| title | Mycobacterium tuberculosis complex detected by modified fluorescent in situ hybridization in lymph nodes of clinical samples |
| title_full | Mycobacterium tuberculosis complex detected by modified fluorescent in situ hybridization in lymph nodes of clinical samples |
| title_fullStr | Mycobacterium tuberculosis complex detected by modified fluorescent in situ hybridization in lymph nodes of clinical samples |
| title_full_unstemmed | Mycobacterium tuberculosis complex detected by modified fluorescent in situ hybridization in lymph nodes of clinical samples |
| title_short | Mycobacterium tuberculosis complex detected by modified fluorescent in situ hybridization in lymph nodes of clinical samples |
| title_sort | mycobacterium tuberculosis complex detected by modified fluorescent in situ hybridization in lymph nodes of clinical samples |
| topic | fluorescent in-situ hybridization FISH tuberculosis Mycobacterium tuberculosis complex PCR insertion sequence 6110 |
| url | https://jidc.org/index.php/journal/article/view/1752 |
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