Fig 1.
Diagram of latex agglutination.
(A) Reaction between gp43-sensitized latex particles (SLPs) for detection of specific antibodies and (B) between the monoclonal antibody mAb17c-SLPs for detection of circulating gp43 antigen in the sample.
Fig 2.
Appearance of typical latex agglutination test.
In positive reactions, the LA tests formed loose aggregates within 5 to 10 min (left). In negative reactions, the LA test remained as a milky suspension (right).
Fig 3.
(A) Detection of anti-gp43 antibody in the sera of patients using gp43-SLPs and (B) gp43 antigen in the sera of patients using mAb17c-SLPs.
Samples were from paracoccidioidomycosis (PCM; n = 65), histoplasmosis (HIS; n = 18), aspergillosis (ASP; n = 18), candidiasis (CAN; n = 13), no fungal disease (NFD; n = 12), and healthy controls (NHS; n = 38). Agglutination score: (4+) 100%, (3+) 75%, (2+) 50%, (1+) 25%, and (N) negative.
Table 1.
Detection of anti-gp43 antibody and gp43 antigen in sera from patients with or without fungal disease.
Fig 4.
(A) Detection of anti-gp43 antibody in the cerebrospinal fluid of PCM patients using gp43-SLPs and (B) gp43 antigen in the cerebrospinal fluid of PCM patients using mAb17c-SLPs.
Agglutination score: (4+) 100%, (3+) 75%, (2+) 50%, (1+) 25%, and (N) negative. Samples were from 14 PCM patients and 6 negative controls with no fungal disease (NFD).
Table 2.
Detection of anti-gp43 antibody and gp43 antigen in cerebrospinal fluid (CSF) from patients with or without paracoccidioidomycosis (PCM).
Fig 5.
(A) Detection of anti-gp43 antibody in the bronchoalveolar lavage fluid from PCM patients using gp43-SLPs and (B) gp43 antigen in the bronchoalveolar lavage fluid from PCM patients using mAb17c-SLPs.
Agglutination score: (4+) 100%, (3+) 75%, (2+) 50%, (1+) 25%, and (N) negative. Samples were from 13 PCM patients and 6 negative controls with no fungal disease (NFD).
Table 3.
Detection of anti-gp43 antibody and gp43 antigen in bronchoalveolar lavage (BAL) fluid from patients with or without paracoccidioidomycosis (PCM).
Fig 6.
Representative curves for serological follow-up.
Samples were from 10 PCM patients with acute (n = 5) or chronic (n = 5) forms of the disease before (T1, at the moment of disease diagnosis), during (T2 = 3 months), and after treatment (T3, between 18–24 months). (A, C) Anti-gp43 antibody detection using gp43-SLPs and (B, D) gp43 antigen detection using mAb17c-SLPs.
Fig 7.
Receiver operating characteristics (ROCs) depicting assay sensitivity and specificity.
Samples were from 65 PCM patient sera and 99 control sera, including patients with histoplasmosis, aspergillosis, non-fungal diseases, and healthy subjects. The ROC is plotted between the true-positive rate (sensitivity) on the y-axis and the false-positive rate (100-specificity) on the x-axis. The area under the curve (AUC) represents the accuracy of the LA test, which was 0.962±0.0143 (95% CI 0.920–0.986, P<0.0001) for the gp43-SLPs, 0.929±0.0192 (95% CI 0.878–0.963, P<0.0001) for the mAb17c-SLPs, and 0.992±0.00769 (95% CI 0.964–1.000, P<0.0001) for the DID. The more the AUC is greater than 0.5, the better the test.