Histopathology. predictive value of 96%. The specificities of the tests are high since none of the 85 control sera was positive by Atosiban Acetate either test. Disseminated systemic fungal infections frequently occur in immunocompromised patients (22). Since many of them fail to produce sufficient levels of specific antibodies that can be recognized by serological checks (22), the analysis of systemic fungal infections often depends on the detection of fungal products or fungal antigens. Analysis of fungal cell wall components revealed the presence of glucan, chitin, and mannoproteins. Cell wall mannan and mannoproteins are abundant and important fungal antigens that represent up to 25% of the total cell wall mass (9, 10, 15). Mannoproteins can be solubilized and removed from the cell surface by denaturation (19) or with reducing providers (13). In addition, the detection of mannan or galactomannan was shown to be useful in the analysis of systemic fungal infections such as 5-Iodo-A-85380 2HCl systemic candidiasis (5, 8) and systemic aspergillosis (14, 21) from the detection of antigenemia. In the study explained here, we explored the possibility of detecting specific mannoproteins in the circulating blood as an alternative approach to the molecular analysis of systemic fungal infections. Previous protein sequence analysis of fungal mannoproteins exposed the presence of a secretory transmission peptide in them (20). Because they are located on the fungal cell wall and may readily be removed from the cells, we rationalized that they may also become secreted from your cells as the fungi grow and divide. The detection of a specific mannoprotein for the analysis of a systemic fungal illness from the detection of antigenemia may have particular advantages. The preparation of the antigen can be more reproducible since it depends on the purification of a specific recombinant protein. In addition, an enzyme-linked immunosorbent assay (ELISA) can be developed with antibodies to a specific mannoprotein. Such an ELISA is better defined and may consequently be more quantitative, sensitive, and specific. is an important dimorphic pathogenic fungus that is endemic in Southeast Asia and southern parts of China. It causes a disseminated and progressive disease, penicilliosis marneffei. The disease happens primarily in AIDS individuals, although it has also been reported to occur in immunocompetent individuals (6, 7, 16, 17, 24). A gene, cells (11), indicating the presence of fungal antigens in the sera of individuals who have been antibody test negative. The present study reports on an ELISA-based test for antigenemia that detects the Mp1p mannoprotein of for the serological analysis of penicilliosis. First, mannoprotein Mp1p was recognized in the cell tradition supernatant of by Western blotting. The anti-Mp1p antibody is definitely specific for since no cross-reaction was observed between cell lysates of and the specific antibodies used to set up the Mp1p ELISA. Next, a sensitive and quantitative ELISA-based antigen test was developed to detect the presence of Mp1p and to quantitate the amount of Mp1p in the cell tradition supernatants of since the cell tradition supernatants of were all negative from the ELISA. Lastly, this Mp1p antigen test matches an ELISA-based Mp1p antibody test for the analysis of systemic penicilliosis. MATERIALS AND METHODS Strains and growth conditions. PM4 and NGY10 were used previously (1, 2). ATCC 26032 was 5-Iodo-A-85380 2HCl from the American Type Tradition 5-Iodo-A-85380 2HCl Collection (Rockville, Md.), and is a medical isolate (from patient 96M0112693) from Queen Elizabeth Hospital, Hong Kong. Fungal cells were first cultivated on YPD plates (1% candida draw out, 2% Bacto Peptone, 2% glucose, 1% agar) at 30C. Fungal cultures were acquired by inoculating fungal cells from plates into RPMI medium (Gibco-BRL, Gaithersburg, Md.) and were further shaken at 37C for 1 to 3 days. Human and animal sera. Sera were obtained from individuals with penicilliosis that was recorded by examination of bone marrow, spleen, pores and skin, or lymph node biopsy specimens and/or blood tradition results..