Brief Article
Copyright ©2012 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Dec 21, 2012; 18(47): 6974-6980
Published online Dec 21, 2012. doi: 10.3748/wjg.v18.i47.6974
Diagnosis of intestinal tuberculosis using a monoclonal antibody to Mycobacterium tuberculosis
Yasushi Ihama, Akira Hokama, Kenji Hibiya, Kazuto Kishimoto, Manabu Nakamoto, Tetsuo Hirata, Nagisa Kinjo, Haley L Cash, Futoshi Higa, Masao Tateyama, Fukunori Kinjo, Jiro Fujita
Yasushi Ihama, Akira Hokama, Kenji Hibiya, Kazuto Kishimoto, Tetsuo Hirata, Haley L Cash, Futoshi Higa, Masao Tateyama, Jiro Fujita, Department of Infectious, Respiratory, and Digestive Medicine, Faculty of Medicine, University of the Ryukyus, Okinawa 903-0215, Japan
Manabu Nakamoto, Nagisa Kinjo, Fukunori Kinjo, Department of Endoscopy, Ryukyu University Hospital, Okinawa 903-0215, Japan
Author contributions: Ihama Y designed and performed the research, analyzed data, and wrote the manuscript; Hibiya K contributed to the experimental concept and provided technical support; Ihama Y, Hokama A, Kishimoto K, Nakamoto M, Hirata T, Kinjo N and Kinjo F contributed to the clinical study; Hokama A and Cash HL reviewed the manuscript; Hokama A, Higa F, Tateyama M, Kinjo F and Fujita J established the research plan and supervised the project; all authors read and approved the data and the final version of the manuscript.
Correspondence to: Akira Hokama, MD, PhD, Assistant Professor, Department of Infectious, Respiratory, and Digestive Medicine, Faculty of Medicine, University of the Ryukyus, Okinawa 903-0215, Japan. hokama-a@med.u-ryukyu.ac.jp
Telephone: +81-98-8951144 Fax: +81-98-8951414
Received: April 19, 2012
Revised: July 19, 2012
Accepted: July 28, 2012
Published online: December 21, 2012
Abstract

AIM: To investigate the utility of immunohistochemical (IHC) staining with an antibody to Mycobacterium tuberculosis (M. tuberculosis) for the diagnosis of intestinal tuberculosis (TB).

METHODS: We retrospectively identified 10 patients (4 males and 6 females; mean age = 65.1 ± 13.6 years) with intestinal TB. Clinical characteristics, including age, gender, underlying disease, and symptoms were obtained. Chest radiograph and laboratory tests, including sputum Ziehl-Neelsen (ZN) staining, M. tuberculosis culture, and sputum polymerase chain reaction (PCR) for tubercle bacilli DNA, as well as Tuberculin skin test (TST) and QuantiFERON-TB gold test (QFT), were examined. Colonoscopic records recorded on the basis of Sato’s classification were also reviewed, in addition to data from intestinal biopsies examined for histopathological findings, including hematoxylin and eosin staining, and ZN staining, as well as M. tuberculosis culture, and PCR for tubercle bacilli DNA. For the present study, archived formalin-fixed paraffin-embedded (FFPE) intestinal tissue samples were immunohistochemically stained using a commercially available species-specific monoclonal antibody to the 38-kDa antigen of the M. tuberculosis complex. These sections were also stained with the pan-macrophage marker CD68 antibody.

RESULTS: From the clinical data, we found that no patients were immunocompromised, and that the main symptoms were diarrhea and weight loss. Three patients displayed active pulmonary TB, six patients (60%) had a positive TST, and 4 patients (40%) had a positive QFT. Colonoscopic findings revealed that all patients had type 1 findings (linear ulcers in a circumferential arrangement or linear ulcers arranged circumferentially with mucosa showing multiple nodules), all of which were located in the right hemicolon and/or terminal ileum. Seven patients (70%) had concomitant healed lesions in the ileocecal area. No acid-fast bacilli were detected with ZN staining of the intestinal tissue samples, and both M. tuberculosis culture and PCR for tubercle bacilli DNA were negative in all samples. The histopathological data revealed that tuberculous granulomas were present in 4 cases (40%). IHC staining in archived FFPE samples with anti-M. tuberculosis monoclonal antibody revealed positive findings in 4 patients (40%); the same patients in which granulomas were detected by hematoxylin and eosin staining. M. tuberculosis antigens were found to be mostly intracellular, granular in pattern, and primarily located in the CD68+ macrophages of the granulomas.

CONCLUSION: IHC staining with a monoclonal antibody to M. tuberculosis may be an efficient and simple diagnostic tool in addition to classic examination methods for the diagnosis of intestinal TB.

Keywords: Colonoscopy, Intestinal tuberculosis, Immunohistochemistry, Monoclonal antibody, Mycobacterium tuberculosis