Analysis of clonality in cytologic material using the polymerase chain reaction (PCR).

Immunoglobulin heavy chain (IgH) gene rearrangement analysis was performed on 27 fine needle aspiration (FNA) specimens (13 reactive hyperplasia, 11 B cell non-Hodgkin's lymphoma (B-NHL), one Hodgkin's disease and two suspicious of non-Hodgkin's lymphoma). Satisfactory amplification was achieved in 23/27 cases. A polyclonal pattern was seen in 14 cases (11 reactive hyperplasia, one B-NHL, one suspicious of lymphoma, one Hodgkin's disease). A monoclonal band was seen in nine cases (eight B-NHL, one reactive hyperplasia). Amplification was unsuccessful in four cases. Clonal analysis by PCR-based IgH gene rearrangement analysis can be successfully applied to FNA material and can be useful in diagnosis, but the results must be interpreted in conjunction with morphology and other ancillary information.

Immunoglobulin light chain mRNA detected by in situ hybridisation in diagnostic fine needle aspiration cytology specimens.

AIMS: To demonstrate expression of immunoglobulin light chain mRNA in diagnostic fine needle aspiration (FNA) cytology specimens using an in situ hybridisation (ISH) technique; and to evaluate ISH in a series of reactive lymphoid proliferations and malignant lymphomas. METHODS: Forty diagnostic FNA specimens showing a lymphoid cell population were examined for immunoglobulin light chain mRNA expression using ISH. Aspirates were obtained from lymph node (n = 34), salivary gland (n = 3), subcutaneous tissue, thyroid and breast (n = 1 each). The cases included 20 B cell lymphomas, five cases of Hodgkin's disease and 15 reactive lymphoid proliferations. Comparison with light chain immunoreactivity was made in 36 cases and histological correlation from biopsy material was available in 24. RESULTS: Immunoglobulin light chain restriction was demonstrated in 14 of 20 B cell lymphomas using ISH and in six of 17 B cell lymphomas using immunocytochemistry. A polytypic pattern of light chain expression was observed in four of five cases of Hodgkin's disease with both techniques, and in 12 of 15 and 11 of 14 reactive lymphoid proliferations using ISH and immunocytochemistry, respectively. CONCLUSIONS: The assessment of immunoglobulin light chain expression is a useful adjunct to morphology in the diagnosis of reactive and malignant lymphoid proliferations in FNA specimens. Light chain restriction can be shown using either immunocytochemistry or ISH, but the latter is more sensitive in the diagnosis of B cell lymphoma.

New method for preserving cytology specimens.

A method of freezing cytological material for long term storage, based on a modification of that used in tissue typing for the storage of lymphocytes, was developed. The method entails the centrifugation of the specimen, adding 1 ml AB or fetal calf serum and 1 ml 20% dimethyl sulphoxide aliquoted into 2 ml tubes, and storage at -70 degrees C. Cytological detail was well preserved in a variety of samples including fine needle aspirates, urine, and bronchial brushings. Three dimensional architecture was also preserved. The method is practical, easy to perform, and allows retrospective studies to be undertaken with the subsequent use of a variety of special staining techniques, including immunocytochemistry.

Parathyroid exploration for primary hyperparathyroidism.

Parathyroidectomy was studied retrospectively in 107 patients with primary hyperparathyroidism. This condition was diagnosed by measuring both the total serum calcium and ultrafilterable calcium (non-protein-bound) levels. The identification of ultrafilterable calcium is an important adjunct to parathyroid surgery as it allows the diagnosis of hyperparathyroidism when the total serum calcium level is normal. The surgical technique for selective parathyroidectomy and multiple biopsies was uniform. Parathyroid adenoma was discovered in 73 patients, diffuse hyperplasia in 26 and combined disease in 8. Postoperatively, two patients suffered from permanent hypocalcemia and three had hypercalcemia.