
(Vol.83 No.7 July 2008)

<1>Kekkaku Vol.83 No.7: 487-496, 2008

Original Article

IDENTIFICATION OF 23 MYCOBACTERIAL SPECIES BY INVADER ASSAY
WITH TARGETING 16S rRNA GENE AND ITS-1 REGION
─ Comparison with DDH Method in Clinical Isolates─

Makoto NAGANO, Sadahiro ICHIMURA, Nobuko ITO, Takayuki TOMII,
Yuko KAZUMI, Katsuaki TAKEI, Chiyoji ABE, and Isamu SUGAWARA

Abstract
　[Purpose and Method] The Invader assay was developed to identify 23 mycobacterial 
species using probes derived from the species-specific region of the 16S rRNA gene 
and the 16S _ 23S rRNA internal transcribed spacer 1 (ITS-1) region, with minor modifications 
of our previous study. In the present study, we compared the identification capability between 
the Invader assay and DNA-DNA hybridization (DDH) method. DDH is commonly used to identify non-
tuberculosis mycobacterium in Japan and 636 clinical mycobacterial strains cultured  on Ogawa 
slants were tested.
　[Results] The Invader assay could identify 615 (96.7％) of the 636 strains. The results con-
tained 14 M.lentiflavum, 3 M.parascrofulaceum and 1 M.intermedium, which were undetectable with 
DDH method. On the other hand, DDH method could identify 580 (91.2％) strains with duplicate 
assay. Of 628 strains except 8 strains identified as a few species by Invader assay, 551 (87.7％)
strains were identified as the same species by two methods. Discordant results were mainly re-
cognized for the identification of M.gordonae, M. avium, M.lentiflavum and M.intracellurare. The 
results of other methods targeting 16S rRNA indicated correctness of the Invader assay.
　[Conclusion] These results indicate that Invader assay could identify more correctly 
than DDH method and could identify about 97％ of clinically important mycobacterium.

Key words: Identification of mycobacteria, Invader assay,16S rRNA gene, ITS-1 region, 
DDH method

Development of Clinical Genomics, BML, Inc., Department of Microbiology, BML, Inc., Mycobacterium 
Reference Center, Research Institute of Tuberculosis, JATA, BML, Inc., Nippon Becton Dickinson 
Company, Ltd.

Correspondence to : Makoto Nagano, Development of Clinical Genomics, BML, Inc., 1361_1, Matoba, 
Kawagoe-shi, Saitama 350 _1101 Japan. 
(E-mail : nagano-m＠bml.co.jp)


<2>Kekkaku Vol.83 No.7: 497-501, 2008

Original Article

EPIDEMIOLOGICAL SURVEY OF HYPERURICEMIA
AS AN ADVERSE REACTION TO ANTITUBERCULOUS THERAPY WITH PYRAZINAMIDE

Hisashi TAKI, Kenji OGAWA, Tatsuya MURAKAMI, and Toshiaki NIKAI

Abstract
　[Purpose] Pyrazinamide is an antituberculous drug that is administered as a two-
month course during treatment of pulmonary tuberculosis. Adverse reactions to 
pyrazinamide have been reported to include hyperuricemia. We performed a retrospective multicenter 
epidemiological survey to assess the relationship between various patient characteristics and the 
uric acid level, the changes of uric acid during pyrazinamide administration, and the use of med-
ications for uric acid control as well as attacks of gout or arthralgia at the onset of hyperuricemia. 
A total of 226 patients who were admitted to four hospitals with pulmonary tuberculosis between January
 and December 2006 and received short-term intensive pyrazinamide therapy were studied.
 [Results] There were 172 men and 54 women with an average age of 59.5 years and an average body mass 
index of 19.8 kg/m2. The average serum uric acid concentration before pyrazinamide treatment was 4.73±
1.78 mg/dl, while the average uric acid level after pyrazinamide treatment was 10.63±2.67 mg/dl, which 
was significantly higher than the pretreatment level (p＜0.0001). During treatment, hyperuricemia 
(Serum uric acid ≧8 mg/dl) was reported in 84.5％ of patients and arthralgia developed in 4.42％. 
Although the therapy instituted in 51 patients (22.57％) had to be interrupted or discontinued 
due to liver dysfunction and skin rashes, which were probably caused by isoniazid and rifampicin, no 
patient ceased taking pyrazinamide due to an increase of uric acid. 
Drugs for uric acid control were administered to 21 patients (9.29％). Pyrazinamide is an important 
agent for intensive short-term antituberculous therapy. Hyperuricemia due to this drug can be managed 
by observation and does not require interruption of dminist-ration.

Key words: Pyrazinamide, Pulmonary tuberculosis, Hyperuricemia, Short-term intensive therapy, Gout

Pharmacy Department, Department of Pulmonary Medicine, National Hospital Organization (NHO) Higashi
Nagoya National Hospital, Department of Microbiology, Pharmacy Faculty, Meijo University

Correspondence to : Hisashi Taki, NHO Higashi Nagoya National Hospital, 5_101, Umemorizaka, Meito-ku, 
Nagoya-shi, Aichi 465 _8620 Japan. 
(E-mail : takih＠toumei.hosp.go.jp)


<3>Kekkaku Vol.83 No.7: 503-506, 2008

Original Article

ESTIMATION OF THE PREVALENCE OF TUBERCULOSIS INFECTION IN THE 1950s IN TOKYO ,

Hirotoshi WATASE, Rika KOIKE, and Chiyo INOGUCHI

Abstract
　[Purpose] We estimated the prevalence of tuberculosis infection in the 1950s in the
urban area of Tokyo using results of the tuberculin skin test (TST) in infants and children.
　[Subject and methods] We analyzed prevalence of tuberculosis using the results of the 
TST in 728 children (5 m.o._24 m.o.) without BCG vaccination in 1954 in Koto Ward, Tokyo.
　[Results] Assuming that the sensitivity and specificity of the TST were 95％ and 98％, respectively, 
the prevalence of TB was estimated to be 2％ (95％ C.I., 0.4_4.3％) among 448 infants under 12 m.o. 
(mean age 0.69 y.o.), and 16％ (95％ C.I., 11.9_21.5％) among 280 children aged 12 m.o._24 m.o. 
(mean age 1.44 y.o.).
　[Conclusion] Being different from the current situation of tuberculosis, the risk of infection
 among infants was high in 1950s in Japan, therefore, it was considered that the prevalence of tuber-
culosis infection rose rapidly among infants in their early period after birth. 
Also, the risk of infection in urban areas was higher comparing with the avrage national rate at that 
time.

Key words : Tuberculosis, Children, Prevalence, Annual risk of infection, Bayes formula

Fukagawa Health Consultation Bureau, Joto South Health Consultation Bureau, Joto Health Consultation 
Bureau, Koto Public Health Center

Correspondence to : Hirotoshi Watase, Fukagawa Health Consultation Bureau, 3_4_3_301, Shirakawa, 
Koto-ku, Tokyo 135_ 0021 Japan. 
(E-mail : h-watase05＠city.koto.lg.jp)


<4>Kekkaku Vol.83 No.7: 507-512, 2008

Original Article

CLUSTERED SECONDARY CASE RATE IN 10,088 PATIENTS WITH TUBERCULOSIS 

Takeo INOUE, Haruki KOYASU, and Satoru HATTORI

Abstract　

 [Objectives] To elucidate characteristics in clustered secondary TB patients trans-
mitted from culture positive pulmonary TB patients.
　[Subjects and Methods] The subjects of this retrospective study were 10,088 TB patients 
registered in Aichi Prefecture between 1989 and 2003. Pulmonary TB was found in 8,629
patients, and 1,459 had extra-pulmonary TB. Bacteriological examination revealed sputum 
smear-positive (SPBP) in 3,332, sputum smear-negative bacillus-culture-positive (SNBP) in
2,139, and smear-negative bacillus-culture-negative (SNBN) in 3,158.
　All registration files were reviewed to identify epidemiological links of patients. When 
linked patients with an interval of the dates of registration of less than 10 years were 
found, the first case was considered as the index case, and the other patients were regarded 
as secondary cases.
　A clustered secondary case rate (CSR) for a category of patients was defined as follows ; 
CSR＝NCS/NA, where NA: number of TB patients in a category A, and NCS: number of secondary 
cases in category A.
　A cluster rate for a category of bacillary pulmonary patients was defined as follows ; 
Cluster rate＝ (NIC＋NCS) /NA, where NA: number of TB patients in a category A, NIC: number 
of index cases in category A, and NCS: number of secondary cases in category A.
　[Results] A total of 417 patients were considered as clustered secondary cases, and the
 CSR was 4.1％ in total. The CSRs were 3.5％ for the SPBP patients, 3.8％ for the SNBP pa-
tients, 5.4％ for the SNBN patients, and 3.4％ for the extra-pulmonary patients. The CSR in 
SNBN patients was significantly higher than the SPBP patients (p＜0.001).
The significant differences in the CSRs were found between the SNBN patients and the SNBP 
patients (p＜0.01), as well as between the SNBN patients and the extra-pulmonary patients 
(p＜0.01).
　The CSRs were 42.5％ in patients aged 0_9, 30.3％ in those aged 10_19, 11.2％ in those 
aged 20_29, 7.4％ in those aged 30_ 9, 4.6％ in those aged 40_49, 3.2％ in those aged 50 
_59, 2.4％ in those aged 60_69, 1.8％ in those aged 70_79, 1.3％ in those aged 80_89, 
and 0.6％ in those aged 90_99. There were significant differences in the CSRs between those 
aged 10_19 and those aged 20_29 (p＜0.001), between those aged 20_29 and those aged 30_ 
39 (p＜0.05), and between those aged 30_39 and those aged 40_49 (p＜0.05). The male pa-
tients showed significantly lower CSR than female patients (2.9％ vs 6.3％, p＜0.001). The 
cluster rate for the 5,471 bacillary patients was 8.8％. The cluster rates were significantly 
different between those patients aged 10 _19 and those aged 20 _ 29 (37.1％ vs 21.1％, 
p＜0.001), as well as between those aged 40_49 and those aged 50_59 (16.4％ vs 8.5％, p＜0.001).

　[Conclusion] These findings suggest that the CSR is closely related with patient’s age, gen-
der, and bacillary findings, and that the CSR is significantly high in young, female, and SNBN
patients.

Key words : Clustered secondary case rate, Cluster rate, Smear-positive pulmonary TB, Culture 
positive pulmonary TB, Close contact, TB transmission, Aging, Gender difference 

Aichi Shikatsu Health Center, Aichi Ichinomiya Health Center, Aichi Toyokawa Health Center

Correspondence to : Takeo Inoue, Aichi Shikatsu Health Center, 114 Shikata Nishimuramae, 
Kitanagoya-shi, Aichi 481_0004 Japan. 
(E-mail : takeo_inoue＠pref.aichi.lg.jp)


<5>Kekkaku Vol.83 No.7: 513-517, 2008

Case Report

A CASE OF MULTI-DRUG RESISTANT PULMONARY TUBERCULOSIS AFTER ADMINISTRATION OF STANDARD ANTI-
TUBERCULOSIS TREATMENT FOR TWO TIMES 

Kiryo WAKABAYASHI, Shuichi YANO, Kanako KOBAYASHI, Yoshiyuki TOKUDA,
Toshikazu IKEDA, Shigenori ISHIKAWA, and Hiroyasu TAKEYAMA

Abstract
　
 We reported a case in which multi-drug resistant tuberculosis was recognized after two
courses of anti-tuberculosis treatment. A 41-year-old woman who had received two courses of anti-
tuberculosis treatment for pulmonary tuberculosis was admitted to our hospital due to productive 
cough, high fever and positive sputum smear showing acid fast bacillus.
In the past treatment, drug susceptibility was unknown because of culture-negative TB. Chest ra-
diograph showed atelectasis of the right upper lobe. The pathological examination of surgically 
resected lung specimen revealed that atelectasis was formed by a granulation tissue with caseous
necrosis progressed to the bronchus wall. We examined cultures three times using both solid and 
liquid media. Liquid culture of the first time specimen was positive for Mycobacterium tuber-
culosis after six weeks and multi-drug resistant tuberculosis was recognized on drug suscepti-
bility test.
Thereafter she was treated with KM, LVFX, PZA and PAS, and maintained sputum smear negative for 
7 months after treatment. Physicians must consider possibility of MDR-TB despite findings showing
smear-positive and culture-negative TB.

Key words : Tuberculosis recurrence, Multi-drug resistant tuberculosis, Smear-positive and culture-
negative TB, TB liquid culture 

Department of Pulmonary Medicine, National Hospital Organization
Matsue National Hospital

Correspondence to : Kiryo Wakabayashi, Department of Pulmonary Medicine, National Hospital 
Organization Matsue National Hospital, 5_8_31, Agenogi, Matsue-shi, Shimane 690_8556 Japan. 
(E-mail : kiryo731＠matsue.hosp.go.jp)


<6>Kekkaku Vol.83 No.7: 519-524, 2008

A CASE OF MILIARY TUBERCULOSIS WITH INTERSTITIAL NEPHRITIS DUE TO RE-ADMINISTRATION OF RIFAMPICIN
TREATED SUCCESSFULLY WITH STEROID 

Takeshi KAWASAKI, Yuka SASAKI, Aya DAN, Rei BEKKU,Tomohiro HASHIMOTO, Masakatsu TAMAKI, 
and Fumio YAMAGISHI

Abstract　

 A 26-year-old man was admitted to a hospital complaing of continuous high fever and ab-
dominal swelling.
As his sputum and ascites culture was positive for acid-fast bacilli and PCR-TB, he was diagnosed 
as miliary tuberculosis, tuberculous with pleuritis and peritonitis, and transferrd to our hospital. 
After initiation of treatment with isoniazid, rifampicin (RFP), ethambutol, and pyrazinamide, RFP 
was suspended because of direct-reacting hyperbilirubinemia. As the liver function recovered after 
discontinuation of RFP, low dose of RFP was re-administrated and renal dysfunction was observed. 
The renal dysfunction continued after discontinuation of suspicious drugs including RFP. As renal 
biopsy revealed interstitial nephritis, prednisolon 20 mg/day was started and renal function re-
covered quickly. From the clinical course and examination, we considered interstitial nephritis
was due to re-administration of RFP and steroid therapy was effective.

Key words : Miliary tuberculosis, Rifampicin, Re-administration,Drug-induced interstitial nephritis, 
Steroid therapy 

Department of Thoracic Disease, National Hospital Organization Chiba-East National 
Hospital

Correspondence to : Takeshi Kawasaki, Department of Thoracic Disease, National Hospital Organization 
Chiba-East National Hospital, 673 Nitona-cho, Chuo-ku, Chiba-shi, Chiba
260_8712 Japan. 
(E-mail : t-kawa＠cehpnet.com.)