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CMV-DNA detection in parenchymatous organs in cases of SIDS

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Abstract

A nested PCR approach has been developed especially for the detection of small amounts of cytomegalovirus (CMV) DNA in autopsy samples. Lung tissue and submandibular glands in 118 cases of infant death (92 SIDS cases, 13 natural deaths due to other defined causes and 13 unnatural deaths) were investigated by this technique and compared to the results obtained by other CMV detection methods (histology, immunohistochemistry, in situ hybridization and PCR). CMV-DNA could be detected in the lung tissue in 7 cases of SIDS using nested PCR. Compared to conventional PCR (3 positive cases in lung tissue) the nested approach always gave glear results and showed less additional bands. In all cases where CMV could be detected in the lungs, positive results were also obtained in the submandibular glands. The nested PCR method proved to be a more sensitive technique than the other detection methods including PCR and hot start, and. even minimal amounts of target DNA could be detected in the presence of human and bacterial background DNA.

Zusammenfassung

Es wurde eine nested PCR Methode speziell für den Nachweis geringer Mengen von Zytomegalievirus DNA in Autopsiematerial entwickelt. In 118 plötzlichen Todesfällen von Säuglingen und Kleinkindern (92 SIDS-Fälle, 13 natürliche Todesfälle anderer definierter Ursache, 13 nichtnatürliche Todesfälle) wurde Lungengewebe und Gewebe der Glandula submandibularis mittels dieser Technik untersucht und mit den Ergebnissen herkömmlicher Methoden (Histologie, Immunhistochemie, in situ Hybridisierung, PCR) verglichen. CMV-DNA konnte dabei durch nested PCR in 7 Fällen im Lungenparenchym nachgewiesen werden. Die “konventionelle” PCR (einschließlich hol Start) ergab in 3 Fällen teils schwach positive Ergebnisse. Beim Vergleich beider Methoden zeigte die nested PCR klarere Ergebnisse und weniger Zusatzbanden. In allen Fällen, in denen in der Lunge positive Ergebnisse erzielt wurden, konnte auch in den Speicheldrüsen CMV DNA nachgewiesen werden. Die nested PCR ist demzufolge verglichen mit PCR und hot start die sensitivste der Methoden und gerade zum Nachweis geringer Mengen von target DNA in Gegenwart von humaner und bakterieller Background DNA geeignet. Die Methode kann sowohl für Formalinfixiertes, paraffineingebettetes wie auch für tiefgefrorenes Material angewendet werden. Sie ist auch zum Virusnachweis in anderen parenchymatösen Organen (Leber, Nieren) geeignet.

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Authors and Affiliations

  1. Institute of Legal Medicine, Westfälische Wilhelms-Universität, Von-Esmarch-Strasse 86, D-48149, Münster, Germany

    Rosanna Cecchi, Thomas Bajanowski & Peter Wiegand

  2. Institute of Microbiology, Westfälische Wilhelms-Universität, Domagkstrasse 10, D-48149, Münster, Germany

    Barbara Kahl

  3. Institute of Legal Medicine, University of Rome, Viale Regina Elena 336, I-00161, Rome, Italy

    Rosanna Cecchi

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  1. Rosanna Cecchi
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  2. Thomas Bajanowski
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  3. Barbara Kahl
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  4. Peter Wiegand
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Cecchi, R., Bajanowski, T., Kahl, B. et al. CMV-DNA detection in parenchymatous organs in cases of SIDS. Int J Leg Med 107, 291–295 (1995). https://doi.org/10.1007/BF01246875

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  • DOI: https://doi.org/10.1007/BF01246875

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