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Taking samples of bone marrow and lymph node for polymerase chain reaction method, immunophenotyping of cells and serving of molecular analysis

Aspirational cytodiagnostic puncture of bone marrow (BM) and the lymph node (LN) as part of the haemathological evaluation process is usually performed by a thin needle pricked into a fixed part of the organ and aspirated via negative pressure to gain samples containing cells. Aim of the study: The purpose of this task is to represent the need for a necessary approach according to regulations on bone marrow and lymph node puncture as the only way to present quantity and quality of samples taken. Although this indicates a small possibility of false positive or negative search results, inadequate puncture or the existence of careless sample manipulation would obstruct or totally disable the analysis. Patients and Methods: At the Laboratory of Cytology and Haematology at the Merkur University Hospital aspiration is used to obtain a cytological analysis that generalizes the morphologic significance of the cells and their disposition, apart from which the above mentioned samples are also taken for certain other diagnostic techniques, such as DNA replication – so called polymerase chain reaction (PCR), immunophenotyping of blood cells by flow cytometry (IF) and cytogenetic methods (karyogram, FISH). Results: During an obseravation period summation was done by 3950 punctures of bone marrow and lymph nodes. In full review of the mentioned aspirations 1526 (38, 63%), as a necessity, specimens were taken at other laboratories. For the PCR analysis 21, 63% of assays (LN 35, BM 295) were aspirated, for the cytogenetic method the samples taken comprise 17, 56% (LN 3, BM 265), while 928 (60, 81%) samples were aspirated for the immunophenotyping method (LN 305, BM 623), from among the aspirations completed collectively. Depending on the sampling method, there was a high degree of sample adequasy. All specimens taken for the PCR method were adequate (100%), and results obtained via the karyogram were also extremely satisfying. Only two samples have not been able to be processed because the culture was insufficient for analysis. In both cases it were bone marrow samples. 18% of the lymph node aspiration samples were unsatisfactory, whereas bone marrow samples were 100% usable. Conclusion: This tests can be used as a supplement to cytological analysis for the purpose of setting an accurate diagnosis. Cytological samples of bone marrow and lymph nodes reveal they are axceptionally adequate for all applied techniques.

samples ; bone marrow ; lymph node ; polymerase chain reaction ; immunophenotyping ; molecular analysis ; cytology

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