／【Ｋ：】Ｓｅｒｖｅｒ／１７０１０３６２ 新潟厚生連農業協同組合／本文／１７ｄ

五十嵐

０８５‐０８８2017.02.10 19.44

A case of Whipple disease diagnosed by
厚生連医誌
polymerase chain
第２６巻
reaction
１号analysis（PCR）
８５〜８８ ２０１７of formalin-fixed paraffin-embedded tissues（FFPE）from duodenum

Brief report

A case of Whipple disease diagnosed by polymerase chain
reaction analysis（PCR）of formalin-fixed paraffin-embeddedtissues（FFPE）from duodenum
Nagaoka Central General Hospital, Department of pathology ; Pathologist

Toshihiko Ikarashi
Background : It was very difficult to diagnose Whippledisease caused by the bacterium Tropheryma whipplei reliably. We tried to confirm this bacterium by
a genetic analysis of histopahtolgical specimen.
Case report : ７
６ year-old male with symptoms of diarrhea, abdominal pain, and weight loss was examined with gastrointestinal endoscope, which revealed atrophic gastritis and white granular appearance in the whole distal duodenum, suggesting
lymphoid hyperplasia. Microhistopathology revealed marked macrophage infiltrate in duodenum,suggesting Whipple disease. He was diagnosed as
Whipple diseases on the basis of the confirmationof Trpheryma whipplei by PCR analysis.
Conclusion : Genetic analysis, especially real-time
method, was very useful and convenient tool forthe diagnosis of Whipple disease.

cobaster pylori and Whipple disease were suggested histologically, the latter of which was speculated because of
the massive histiocytic infiltrate immunohistochemicallyand the intracellular digestion-resistant granules on digested Periodic acid-Schiff stain（Fig. １-３）
. Final diagnosis of Whipple disease was confirmed by the following
genetic studies :（i）uPCR with EP,（ii）real-time PCR.
Deoxyribonucleic acid（DNA）extracts were obtained by
the previous method（１-３）. The condition of PCR inuPCR with EP was same as the report（１,２）. Tropheryma
whipplei１６S rDNA gene from９９５to１２６０ was amplified
with the primers, produced the final product of ２６６ bps（４）. The concrete technique of Real-time PCR was described in our previous report（３）. The specific dissociation peak temperature was ８９．
６８℃. The technical simplicity and its required time in Real-time PCR method
was more convenient than those in uPCR with EP.

Key words : Whipple disease, duodenum, polymerase
chain reaction（PCR）, PCR, real-time SYBR Green
method, formalin-fixed paraffin-embedded tissues（FFPE）, immunohistochemistry

Conclusion
Genetic analysis, especially real-time method, wasvery useful and convenient tool for the diagnosis of
Whipple disease.

Background
Reference

Whipple disease was a rare systemic infectious
disease caused by the bacterium Tropheryma whipplei,commonly involved duodenum with malabsorption. This
life-threatening disease could be treatable with long-term
antibiotic therapy, which required the reliable diagnosticmethod. Genetic confirmation of this bacterium was expected as one of the most reliable diagnostic tools. TheFFPE specimens of duodenum was studied genetically byboth the usual PCR followed electrophoresis（uPCR withEP）and the real-time SYBR Green method（real-timePCR）, the superiority of which was compared.

１．Ikarashi, T, Hasegawa H. Detection of monoclonality
in B- and T-cell lymphoma by the use of polymerase
chain reaction of formalin-fixed paraffin-embedded tissues. Niigata-ken Koseiren Med J２０００;１０:１０−５．
２．Ikarashi, T, Hasegawa H. Improved detection ofmonoclonality in B- and T-cell lymphomas by polymerase chain reaction（PCR）of formalin-fixed paraffinembedded tissues with the use of deoxyribonucleic acid（DNA）purification and concentration : Efficacy ofPCR-PAGE of Byori Center method for lymphoma,
００１;１１:２
５−９．
Ver.２.２
３．Ikarashi, T. Trial of rapid diagnosis of infection andindividual confirmation against tissue contamination byreal time PCR（polymerase chain reaction）of SYBRGreen method to paraffin-embedding specimens. Niigata-Ken Koseiren Med J２０１６;２５:１２７−３０．

Case report
７６ year-old male with symptoms of diarrhea, abdominal pain, and weight loss was examined with gastrointestinal endoscope. Chronic atrophic gastritis with Heli-― 85 ―

／【Ｋ：】Ｓｅｒｖｅｒ／１７０１０３６２ 新潟厚生連農業協同組合／本文／１７ｄ

厚生連医誌

第２６巻

五十嵐

０８５‐０８８2017.02.10 19.44

１号 ８５〜８８ ２０１７４．Yajima, N et al. Whipple disease diagnosed with
PCR using formalin-fixed paraffin-embedded specimensof the intestinal mucosa. Intern Med ２０１３;５２:２
１９−２２．

和 文 抄

プル病を確実に診断することは非常に難しい。
我々は、通常病理組織標本の遺伝分析によってこの細菌を確認し、確診できたので報告した。
症例報告：症例は、下痢、腹痛と体重減少を症状とした７６歳の男性で、上部消化管内視鏡検査が施行された。萎縮性慢性胃炎と遠位十二指腸のリンパ球増殖を示唆する白色顆粒状像が認められた。病理組織検査上、十二指腸には組織球の顕著な浸潤が明らかにされ、ウイップル病が示唆された。PCR 分析による Trpheryma whipplei の確認により、ウイップル病と確診された。
結論：遺伝学的分析（特にリアルタイム法）は、ウイップル病の診断において非常に役立ち、かつ、便利な検査法であった。

録短報
十二指腸のホルマリン固定パラファフィン包埋材料（FFPE）を使ったポリメラーゼ連鎖反応遺伝子検査（PCR）によりウイップル病と診断された１例
長岡中央病院、病理部；病理医
いキーワード：ウイップル病、ポリメラーゼ連鎖反応（PCR）、リアルタイム SYBR グリーン方法、
ホルマリン固定パラフィン包埋組織（FFPE）、免疫組織化学か らしとしひこ

五十嵐俊彦
背景：Tropheryma whipplei 細菌感染に起因するウイッFig. １. Marked foamy cell clusters were shown in mucosa
by Hematoxyline-eosin stain.

Fig. ２. Marked histiocytic infiltrate was confirmed by immunostain with CD６８．

― 86 ―

／【Ｋ：】Ｓｅｒｖｅｒ／１７０１０３６２ 新潟厚生連農業協同組合／本文／１７ｄ

五十嵐

０８５‐０８８2017.02.10 19.44

A case of Whipple disease diagnosed by polymerase chain reaction analysis（PCR）
of formalin-fixed paraffin-embedded tissues（FFPE）from duodenum

Fig.３. Positive intracellular granules were found on digested
Periodic acid-Schiff stain（PAS）.

Fig. ４. Single band was found at ２６６ bp on electrophoresis after polymerase chain reaction.

― 87 ―

／【Ｋ：】Ｓｅｒｖｅｒ／１７０１０３６２ 新潟厚生連農業協同組合／本文／１７ｄ

厚生連医誌

第２６巻

五十嵐０８５‐０８８

2017.02.10 19.44

１号 ８５〜８８ ２０１７Fig.５. Amplification elevated at２２cycles on real-time polymerase chain reaction.

Fig.６. Dissociation peak temperature was８９．
６８℃ on real-time polymerase chain reaction.
（２０１６／１２／３０受付）― 88 ―