Recombinant antigen-based immuno-slot blot method for serodiagnosis of syphilis

Three recombinant antigens of Treponema pallidum Nichols strain were fused with GST, cloned and expressed in Escherichia coli, resulting in high levels of GST-rTp47 and GST-rTp17 expression, and supplementation with arginine tRNA for the AGR codon was needed to obtain GST-rTp15 overexpression. Purified fusion protein yields were 1.9, 1.7 and 5.3 mg/l of cell culture for GST-rTp47, GST-rTp17 and GST-rTp15, respectively. The identities of the antigens obtained were confirmed by automated DNA sequencing using ABI Prism 310 and peptide mapping by Finningan LC/MS. These recombinant antigens were evaluated by immuno-slot blot techniques applied to 137 serum samples from patients with a clinical and laboratory diagnosis of syphilis (61 samples), from healthy blood donors (50 samples), individuals with sexually transmitted disease other than syphilis (3 samples), and from individuals with other spirochetal diseases such as Lyme disease (20 samples) and leptospirosis (3 samples). The assay had sensitivity of 95.1 percent (95 percent CI, 86.1 to 98.7 percent) and a specificity of 94.7 percent (95 percent CI, 87.0 to 98.7 percent); a stronger reactivity was observed with fraction rTp17. The immunoreactivity results showed that fusion recombinant antigens based-immuno-slot blot techniques are suitable for use in diagnostic assays for syphilis.

Country reports from Japan: external quality assessment scheme in Japan.

Several external quality assessment schemes (EQAS) have been conducted in Japan. Results obtained from nation-scale EQAS reveal the current quality of laboratory testing in each laboratory. The largest nation-scale EQAS in Japan is that conducted by the Japan Medical Association. The numbers of participants and of items evaluated have increased in EQAS by JMA over its history of 32 years. Improvement in inter-laboratory differences has been observed for most items in EQAS in recent decades. In 1998, about 2,500 laboratories from throughout the country participated in this surveillance, and 47 items were evaluated. The coefficient of variations for the group of all participants was less than 5% for about one third of all test items. On the other hand, very high variations over 20% were observed for 6 items. Also, inter-method differences exist for many items, which may be or may not be related to matrix effects. Retrospective evaluation of all EQAS data suggests that there is still room for improvement in inter-laboratory differences.

External quality control study of lymphocyte subset tests in Japan.

We have evaluated external quality assurance of lymphocytes subset tests in Japan. The data suggest: 1) Values of lymphocyte subsets were variable at each medical laboratory in Japan. 2) Value of CD3 did not varied so much, but that of CD 19/20 or NK marker was widely varied. 3) Values of CD4 and CD8 varied in small extension. 4) CV of values was relatively low when analysis was carried out in commercial laboratories or when antibodies and instrument of Ortho-Clinical Diagnostics were used. 5) In some cases, CV was also relatively low if number of cell washing times was low or if self made lytic agents were used for cell lysing.

Susceptibilidade aos antimicrobianos de Listeria spp isoladas de pacientes com aborto repetitivo / Antibiotic susceptibility testing of Listeria spp from endocervix of women with repeated abortion

Neste estudo foi verificada a susceptibilidade "in vitro" aos antimicrobianos de Listeria spp isoladas de endocévix de mulheres com clínica de aborto repetitivo. O tratamento de listeriose com antibióticos mais frequentemente recomendados (penicilina e ampicilina) é geralmente bem sucedido, porém há vários casos notificados de falhas terapêuticas. Foi utilizado o método de macrodiluiçäo em tubo para determinar as concentraçöes mínimas inibitórias (CMIs). A ampicilina e a penicilina foram bacteriostáticas para as Listeria spp. A gentamicina e a ciprofloxacina foram bactericidas, e CMBs foram até 3 vezes maiores que as respectivas CMIs. Por outro lado, as CMBs de penicilina e ampicilina foram pelo menos 6 vezes maiores que CMIs. Três amostras de Listeria monocytogenes mostraram tolerância para com penicilina e ampicilina

Endocarditis caused by Candida parapsilosis.

The authors report a case of endocarditis caused by Candida parapsilosis. To the best of our knowledge, a case has not been described previously in Japan in the English literature. A battery of 8 peroxidase-labeled lectins was tested on sections of paraffin-embedded tissue to determine which lectin could be used in the microscopic diagnosis of C. parapsilosis. One lectin, from Archis hypoaea (PNA) was found to react with C. parapsilosis. On the other hand, C. albicans, Aspergillus, Mucor, and Cryptococcus did not react with A. hypoaea (PNA). On fluorescence microscopic study, C. parapsilosis was not fluorescent, but other fungi were fluorescent when exposed to ultraviolet illumination. Therefore, we propose new procedures for identification of C. parapsilosis in tissue sections using lectin histochemistry and fluorescence microscopy.