Multiplex STR and mitochondrial DNA testing for paraffin embedded specimen of healthy and malignant tissue: Interpretation issues

Lü, Chan; Budimlija, Zoran; Popiolek, Dorota; Illei, Peter B.; West, Brian K; Fogt, Franz; Wei, Jinlong; Prinz, Mechthild

doi:10.1016/j.ics.2005.10.010

Cited by 4 publications

(4 citation statements)

References 5 publications

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“…In general, histological preparations are formalin‐fixed and paraffin‐embedded tissue samples, which can cause problems in the interpretation of DNA profiles because of degradation processes during fixation (10,11). Preferential amplification and allelic drop‐out caused by degradation processes cannot always be distinguished from LOH in cancerous material and interpretation rules would be useful (12).…”

Section: Resultsmentioning

confidence: 99%

Identification of Biological Samples in a Case of Contamination of a Cytological Slide Preparation*

Junge¹,

Dettmeyer²,

Madea³

2008

Journal of Forensic Sciences

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Here we report a case where a discrete section of the cytological slide preparation of a female individual was obviously contaminated with pleura liquid of a female tumor patient. Analysis of the cancerous pleura liquid and the healthy cells of the slide preparation showed different DNA profiles, indicating that the material originated from two different female individuals. The DNA profile of the cell mixture revealed a heterogenous pattern whereby the alleles could be assigned to the healthy and the tumor patient. Loss of heterozygosity (LOH) was observed in four of eight short tandem repeat systems for the pleura liquid and the cell mixture. Despite the low amount of DNA on the slide preparation and the occurrence of LOH, it was possible to clarify the case and to support the assumption that a drop of cancerous pleura liquid contaminated the cytological slide.

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Section: Resultsmentioning

confidence: 99%

Identification of Biological Samples in a Case of Contamination of a Cytological Slide Preparation*

Junge¹,

Dettmeyer²,

Madea³

2008

Journal of Forensic Sciences

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“…The natural germline mutation rate of STRs is estimated to be between 10 −3 and 10 −4 per generation (15,17), which is low enough to provide the stability and reliability through generations for forensic identification and paternity testing. However, these and other STR markers are proven to be unstable in various tumor tissues (18,19).…”

Section: Genetic Instability Of Tested Tumor Samplesmentioning

confidence: 99%

“…Part of the study reported herein was presented in the form of preliminary data as a poster entitled: "Multiplex STR and Mitochondrial DNA testing for Paraffin Embedded Specimen of Healthy and Malignant Tissues: Interpretation Issues" at the 21st Congress of the International Society of Forensic Genetics (ISFG) held at Ponta Delgada, Azores, Portugal, September [13][14][15][16][17]2005.…”

Section: Disclaimermentioning

confidence: 99%

Malignant Tumors and Forensics – Dilemmas and Proposals

et al. 2009

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Aim To evaluate the effect of genetic instability and degradation in archived histology samples from cancerous tumors and to investigate the validity of short tandem repeat (STR) typing of these samples and its potential effect on human identification.Methods Two hundred and twenty eight slides of archival pathology tissues from 13 different types of malignant tumors were compared with healthy tissues from the same individuals. DNA analysis was performed using standard techniques for forensic STR analysis, PowerPlex®16 and Identifiler® on 2 distinct sample sets. Genetic instability was assessed by comparing reference tissues with cancerous tissues derived from the same individual. Loss of heterozygosity, a ≥50% reduction in heterozygosity ratio between healthy and diseased samples, and microsatellite instability, the presence of an additional allele not present in reference tissue, were assessed. The quality of profiles obtained with respect to completeness among the archived samples and degradation using the 2 platforms were also compared.Results Profiles obtained using the Identifiler® system were generally more complete, but showed 3-fold higher levels of instability (86%) than those obtained using PowerPlex® 16 (27%). Instances of genetic instability were distributed throughout all loci in both multiplex STR systems. ConclusionAfter having compared 2 widely used forensic chemistries, we suggest individual validation of each kit for use with samples likely to exhibit instability combined with fixation induced degradation or artifact. A "one size fits all" approach for interpretation of these samples among commercially available multiplexes is not recommended.

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“…The analysis of mtDNA offers advantages over nuclear DNA due to a larger number of copies per cell, what potentiates the recovery of DNA from difficult or degraded materials (4,7) . The literature describes several successful protocols for analysis of shorter and longer mtDNA fragments recovered from archival fixed paraffinembedded human tissues (1,6,11) . In our series of cases, the success of mtDNA analysis was achieved by the use of a DNA extraction kit with modifications in several steps of the process: lysis (additional lysis step), extraction (reduction of buffer volumes and homogenization by gentle handling), amplification (high-annealing temperature and increase in the number of PCR cycles) and sequencing (adjustment of the volume of the PCR product).…”

Section: Resumo Abstractmentioning

confidence: 99%

Optimization of methods to assess mitochondrial DNA in archival paraffin-embedded tissues from mammary canine tumors

Bertagnolli¹,

Máximo²,

Asch³

et al. 2008

J. Bras. Patol. Med. Lab.

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A presente comunicação descreve as modificações usadas para extrair e amplificar o DNA mitocondrial obtido de amostras de tumores mamários caninos fixados em formol tamponado a 10% e incluídos em parafina. Os componentes epiteliais e mesenquimais (condromixóide e condróide), bem como a mama normal adjacente, foram microdissectados manualmente de 19 tumores mamários (10 tumores mistos benignos e nove carcinomas em tumores mistos). O DNA foi extraído utilizando-se o Invisorb ® Spin Tissue Mini Kit com modificações do protocolo proposto pelo fabricante. Um fragmento de 273-pb foi amplificado por reação em cadeia da polimerase (PCR) e seqüenciado em seqüenciador automático. O fragmento foi analisado em 100% das amostras, entretanto modificações como lise adicional, redução do volume das soluções de extração e PCR, aumento da temperatura de anelamento e do número de ciclos de amplificação foram necessárias. Em seis amostras os produtos iniciais de PCR foram diluídos e reamplificados para obtenção de sucesso. resumo abstractIn this study we describe the alterations used to extract and amplify mitochondrial desoxyribonucleic acid (DNA) from formalin-fixed paraffin-embedded samples of canine mammary tumors. The epithelial and mesenchymal components (chondromyxoid and chondroid) of each tumor, as well as the normal mammary gland tissues, were manually microdissected from 19 mixed canine mammary tumors (10 benign mixed tumors and nine carcinomas arising in mixed tumors). DNA was extracted by Invisorb ® Spin Tissue Mini Kit, with protocol changes proposed by the manufacturer. A 273-bp fragment was amplified by polymerase chain reaction (PCR) and submitted to automatic sequence analysis. The fragment was successfully analyzed in 100% of the samples. However, an additional lysis step, the reduction of volume in buffer solutions and PCR, a higher annealing temperature and an increase in the number of PCR cycles were required. The initial PCR products were diluted and re-amplified in six unitermos key words Neoplasia Cão Mesenquimal Neoplasia Dog Mesenchymal Primeira submissão em 06/05/08 Última submissão em 10/10/08 Aceito para publicação em 09/11/08 Publicado em 20/08/08 J Bras Patol Med Lab • v. 44 • n. 4 • p. 305-308 • agosto 2008

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Multiplex STR and mitochondrial DNA testing for paraffin embedded specimen of healthy and malignant tissue: Interpretation issues

Cited by 4 publications

References 5 publications

Identification of Biological Samples in a Case of Contamination of a Cytological Slide Preparation*

Identification of Biological Samples in a Case of Contamination of a Cytological Slide Preparation*

Malignant Tumors and Forensics – Dilemmas and Proposals

Optimization of methods to assess mitochondrial DNA in archival paraffin-embedded tissues from mammary canine tumors

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