Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions
Introduction: The exposure of nervous tissue samples to high temperatures affects the sensitivity of rabies virus diagnostic tests, causing degradation of the viral structure. This study evaluated reverse transcriptase polymerase chain reaction (RT-PCR) for the diagnosis and molecular characterizati...
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| Format: | Article |
| Language: | English |
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The Journal of Infection in Developing Countries
2014-08-01
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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/4136 |
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| author | Fernando J Beltran Federico Gury Dohmen Horacio Del Pietro Daniel M Cisterna |
| author_facet | Fernando J Beltran Federico Gury Dohmen Horacio Del Pietro Daniel M Cisterna |
| author_sort | Fernando J Beltran |
| collection | DOAJ |
| description | Introduction: The exposure of nervous tissue samples to high temperatures affects the sensitivity of rabies virus diagnostic tests, causing degradation of the viral structure. This study evaluated reverse transcriptase polymerase chain reaction (RT-PCR) for the diagnosis and molecular characterization of brain tissue samples in an advanced state of decomposition and poorly conserved viral isolates by comparing it with routine diagnostic tests.
Methodology: A panel of three canine brain samples exposed to controlled decomposition for 7, 15, 30, and 120 days were evaluated using fluorescence antibody test (FAT), mouse inoculation test (MIT), and RT-PCR. In addition, 14 isolates of rabies variants, representing the largest circulation in Argentina, preserved in inadequate cooling for six to eight years were analyzed. Molecular typing of strains was performed using a 159-nucleotide region corresponding to the nucleoprotein gene.
Results: The three samples analyzed were positive by RT-PCR at all the decomposition times evaluated, in contrast to results observed with FAT and MIT, which rapidly became negative. In addition, 100% of the inadequately preserved samples were characterized molecularly. The limit of detection of RT-PCR was 0.5 MICDL50/0.03 mL.
Conclusion: RT-PCR can be useful for rabies diagnosis and typing of putrefying samples or rabies isolates stored in inadequate conditions.
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| format | Article |
| id | doaj-art-ad12de1f6426499aa92059c6b842f2c9 |
| institution | OA Journals |
| issn | 1972-2680 |
| language | English |
| publishDate | 2014-08-01 |
| publisher | The Journal of Infection in Developing Countries |
| record_format | Article |
| series | Journal of Infection in Developing Countries |
| spelling | doaj-art-ad12de1f6426499aa92059c6b842f2c92025-08-20T02:14:14ZengThe Journal of Infection in Developing CountriesJournal of Infection in Developing Countries1972-26802014-08-0180810.3855/jidc.4136Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditionsFernando J Beltran0Federico Gury Dohmen1Horacio Del Pietro2Daniel M Cisterna3Instituto de Zoonosis Luis Pasteur, Buenos Aires, ArgentinaInstituto de Zoonosis Luis Pasteur, Buenos Aires, ArgentinaServicio Nacional de Sanidad y Calidad Agroalimentaria, Buenos Aires, ArgentinaServicio de Neurovirosis, INEI-ANLIS Dr. Carlos G. Malbrán, Buenos Aires, ArgentinaIntroduction: The exposure of nervous tissue samples to high temperatures affects the sensitivity of rabies virus diagnostic tests, causing degradation of the viral structure. This study evaluated reverse transcriptase polymerase chain reaction (RT-PCR) for the diagnosis and molecular characterization of brain tissue samples in an advanced state of decomposition and poorly conserved viral isolates by comparing it with routine diagnostic tests. Methodology: A panel of three canine brain samples exposed to controlled decomposition for 7, 15, 30, and 120 days were evaluated using fluorescence antibody test (FAT), mouse inoculation test (MIT), and RT-PCR. In addition, 14 isolates of rabies variants, representing the largest circulation in Argentina, preserved in inadequate cooling for six to eight years were analyzed. Molecular typing of strains was performed using a 159-nucleotide region corresponding to the nucleoprotein gene. Results: The three samples analyzed were positive by RT-PCR at all the decomposition times evaluated, in contrast to results observed with FAT and MIT, which rapidly became negative. In addition, 100% of the inadequately preserved samples were characterized molecularly. The limit of detection of RT-PCR was 0.5 MICDL50/0.03 mL. Conclusion: RT-PCR can be useful for rabies diagnosis and typing of putrefying samples or rabies isolates stored in inadequate conditions. https://jidc.org/index.php/journal/article/view/4136rabiesmolecular typingdecomposed samples |
| spellingShingle | Fernando J Beltran Federico Gury Dohmen Horacio Del Pietro Daniel M Cisterna Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions Journal of Infection in Developing Countries rabies molecular typing decomposed samples |
| title | Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions |
| title_full | Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions |
| title_fullStr | Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions |
| title_full_unstemmed | Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions |
| title_short | Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions |
| title_sort | diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions |
| topic | rabies molecular typing decomposed samples |
| url | https://jidc.org/index.php/journal/article/view/4136 |
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