Genetic analysis of hydatidiform moles in paraffin wax embedded tissue using rapid, sequence specific PCR-based HLA class II typing.

Abstract

Aims-To determine the applicability of rapid, sequence specific polymerase chain reaction (PCR)-based HLA class II genotyping for the distinction ofcomplete from partial hydatidiform moles (HM) using DNA extracted from formalin fixed and paraffin wax embedded tissue. Methods-Nine HM were studied. DNA was extracted from formalin fixed and paraffin wax embedded tissue after mechanical separation of decidual and molar components. HLA class II DRB (DRB1, -3, -4, and -5) and DQB1 genotyping was performed using a parallel series of PCR reactions, each of which containeUd sequence specific primers designed to amplify different HLA DRB and DQB1 alleles or allele groups (PCR-SSP analysis). In each case the HLA DRB and DQB1 genotypes identified within the decidua and HM were compared. Results-Within the decidual tissue, HLA DRB genotypes were assignable in all nine cases, and HLA DQBl genotypes were identified in seven cases. Within the molar tissue, HLA DRB genotypes were assignable in seven cases, and at least one HLA DQB1 allele was identified in seven cases. Interpretation based on HLA class II genotyping was therefore possible in two cases classified on histological appearances as complete HM, in four classified as partial HM, and in one HM of uncertain type. Different HLA DRB and DQB1 haplotypes were identified within the decidual and molar tissue from both complete HM, consistent with a solely paternal origin and supporting the histological diagnosis. HLA DRB and DQB1 alleles common to the decidual and molar tissue were present within the four partial HM and the HM of histologically uncertain type, consistent with combined maternal and paternal genetic input to these HM, supporting the histological diagnosis in four cases and suggesting that the histologically equivocal case was also a partial HM.Conclusion-PCR-SSP HLA class II DRB and DQB1 typing is reliably applicable to DNA extracted from formalin fixed and paraffin wax embedded tissue. Therefore, in a suitably equipped HLA typing laboratory, this technique provides a useful adjunct to histological examination for differentiation of complete from partial HM. (7 Clin Pathol 1997;50:288-293)

p57KIP2 immunohistochemistry in early molar pregnancies: emphasis on its complementary role in the differential diagnosis of hydropic abortuses

“…Ploidy studies in these cases are of relatively limited usefulness because both HA and CM are diploid in DNA content. More sophisticated molecular genetic techniques have subsequently been applied to resolve this differential diagnostic dilemma, but these assays are largely outside the realm of most pathology laboratories [10,14,15,17].…”

“…Molar pregnancies are being evacuated early in gestation, before the development of wellestablished classic morphologic features, thus adding to the difficulty in diagnosis. Pathologists now rely on molecular techniques that make use of DNA content differences between CM and PM, including DNA flow or image cytometry, chromosome in situ hybridization, polymerase chain reaction-based genotyping or HLA typing, to help in the differential diagnoses of these hydropic placentas [10,[14][15][16][17]. However, these techniques are technically difficult and relatively expensive, and unlikely to become routine in all laboratories.…”

A novel approach for HLA-A typing in formalin-fixed paraffin-embedded-derived DNA

“…This method was also attempted for HLA-typing our samples with no results, which possibly was due to the age of our samples, which may have accumulated more DNA damage. Furthermore, in two studies from 1997, Bateman et al, 22,23,25 showed that the PCR-SSP approach could generate HLA typing in formalinfixed paraffin-embedded material. In both studies only HLA class II was determined, and in the first only skin tissue was used with conventional PCR-SSP, whereas in the second a variety of tissues were used, but only totally five samples were run with a nested PCR-SSP.…”

“…[22][23][24][25] Recently, eg Ota et al 24 showed successful HLA typing in different types of tissue after 24-96 h of formalin fixation, using a PCR-SSOP method for both HLA class I and II typing. This method was also attempted for HLA-typing our samples with no results, which possibly was due to the age of our samples, which may have accumulated more DNA damage.…”

Analysis of gestational trophoblastic disease by genotyping and chromosome in situ hybridization

“…[10][11][12][13] As a result, techniques that make use of the ploidy difference between complete and partial moles such as DNA flow or image cytometry, 5,[14][15][16][17][18][19][20] chromosome in situ hybridization, 14,15,17 as well as polymerase chain reaction (PCR)-based microsatellite genotyping [21][22][23] and HLA typing 24 have been evolved to help in distinguishing complete from partial moles.…”