2007 Fiscal Year Final Research Report Summary
New method for nucleic acid amplification on pathologic tissue section of malignant lymphoma using LAMP method
| Project/Area Number |
18590341
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Human pathology
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| Research Institution | Nara Medical University |
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Principal Investigator |
NAKAMINE Hirokazu Nara Medical University, Dept Med, Assoc. Prof. (70155810)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHIKO Kasai Nara Medical University, Dept Med, Instructor (50224374)
ENOMOTO Yasunori Nara Medical University, Dept Med, Instructor (60336849)
TAKEDA Maiko Nara Medical University, Dept Med, Instructor (40398441)
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| Project Period (FY) |
2006 – 2007
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| Keywords | Malignant lymphoma / LAMP method / in situ nucleic acid amplification / Follicular lymphoma / bcl-2 rearrangement / FISH method |
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| Research Abstract |
Classification of malignant lymphoma is important for selection of treatment modality as well as for investigation of pathogenesis. Because B-cell lymphomas consist of several types which are correspond well to chromosomal/genetic abnormalities, more accurate classification can be performed when these abnormalities are detectable at daily pathology practice. However ; it is rather general in Japan that the first biopsy for pathological diagnosis is often performed at the regional hospitals rather than at the specialized medical institutions. The patients are transferred to the latter institutions where the second biopsy is sometimes required for examination of chromosomal/genetic abnormalities. This is due to the limitation and difficulty in performing such examination on paraffin-embedded tissue. To overcome this problem, we attempted to establish a new method for microscopic visualization of these abnormalities by amplification of genes involved in them on microscopic glass slides. We employed loop-mediated isothermal amplification (LAMP) instead of conventional PCR for this in situ DNA amplification, because LAMP method has several advantages over the conventional PCR for this purpose, including lower degree of tissue damages by using lower temperature, use of small-sized DNA polymerase, and larger size of amplification products. We fried to in situ amplify the rearranged bcl-2/IgH genes as a result of t (14 ; 18) which are present in more than 80% cases of follicular lymphoma. However, this attempt appears to be more difficult than initially thought and satisfactory amplification has not been obtained so far In the literature, only a single report of successful detection of point mutation of some gene by this in situ LAMP (Ikeda, et. al. Pathol Int 57, 594-599, 2007), indicating that there may be unexpected problems. Through this trial, however ; several minor findings regarding B-cell lymphoma could be obtained.
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