Naslov
Heteroduplex analysis in discrimination between monoclonal and polyclonal PCR products

Autori
Kardum Paro, Mirjana Mariana ; Šiftar, Zoran ; Korać, Petra ; Dominis, Mara ; Flegar- Meštrić, Zlata

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, stručni

Izvornik
18th IFCC-EFCC European Congress of Clinical Chemistry and Laboratory Medicine, Innsbruck, Austria, June 7-11, 2009:W-A178. Clin Chem Lab Med 2009 ; 47, Special Suppl., S305. / - Innsbruck : Walter de Gruyter, 2009

Skup
Euromedlab Innsbruck 2009 ; 18th IFCC-EFCC European Congress of Clinical Chemistry and Laboratory Medicine

Mjesto i datum
Innsbruck, Austrija, 07.06.2009. - 11.06.2009

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
PCR-based clonality; Non- Hodgkin΄s lymphoma; heteroduplex analysis

Sažetak
Background. PCR-based clonality methods are crucial for the detection of clonally rearranged immunoglobulin (Ig) and T-cell receptor (TCR) genes in case of suspected Non- Hodgkin΄s lymphoma (NHLs), but require additional analysis of PCR products for discrimination between monoclonal and polyclonal samples. Although the test manufacturers suggested agarose TBE or polyacrylamide TBE gels detection as one way of detection, according to European BIOMED-2 collaborative study only two techniques are recomended: heteroduplex analysis or GeneScanning. After unexpected PCR amplification gel detection results, we performed PCR amplification followed by heteroduplex analysis. Aim. To compare PCR amplification followed by heteroduplex analysis gel detection and PCR amplification gel detection. Methods. DNA samples taken from 15 patients with suspected NHLs are amplified with multiplex master mixes containing consensus DNA primers that target different regions within Ig/TCR genes (IgH and IdentiClone TCRB Gene Clonality Assay, Ipsogen). PCR amplification followed by heteroduplex analysis gel detection and PCR amplification gel detection were analysed in parallel. Clonal rearrangements were identified as single-sized PCR products inside the valid size range. Results. Final PHD diagnosis of NHL was found in 47% (7/15) of suspected NHLs whereas 40% were reactive (6/15). In 6 cases of 7 NHLs with PCR amplification gel detection clonal rearrangement was found while in only 2 of 7 NHLs clonal rearrangement after PCR amplification followed by heteroduplex analysis gel detection was found. Conclusion. PCR amplification followed by heteroduplex analysis gel detection in cases of suspected NHLs reduces false- positive PCR results and therefore increased the test specificity.

Izvorni jezik
Engleski

Znanstvena područja
Kliničke medicinske znanosti

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