Strongyloides venezuelensis is a parasitic nematode of rodents
frequently used to obtain heterologous antigens for the immunological diagnosis of
human strongyloidiasis. The aim of this study was to evaluate membrane fractions from
S. venezuelensis for human strongyloidiasis immunodiagnosis.
Soluble and membrane fractions were obtained in phosphate saline (SS and SM) and
Tris-HCl (TS and TM) from filariform larvae of S. venezuelensis.
Ninety-two serum samples (n = 92) were obtained from 20 strongyloidiasis patients
(Group I), 32 from patients with other parasitic diseases (Group II), and 40 from
healthy individuals (Group III), and were analyzed by enzyme-linked immunosorbent
assay (ELISA). Soluble fractions (SS and TS) showed 90.0% sensitivity and 88.9%
specificity, whereas the membrane fractions (SM and TM) showed 95.0% sensitivity and
94.4% specificity. The present results suggest the possible use of membrane fractions
of S. venezuelensis as an alternative antigen for human
strongyloidiasis immunodiagnosis.
Rapid diagnostics is pivotal to curb SARS-CoV-2 transmission, and saliva has emerged as a practical alternative to naso/oropharyngeal (NOP) specimens. We aimed to develop a direct RT-LAMP (reverse transcription loop-mediated isothermal amplification) workflow for viral detection in saliva, and to provide more information regarding its potential in curbing COVID-19 transmission. Clinical and contrived specimens were used to optimize formulations and sample processing protocols. Salivary viral load was determined in symptomatic patients to evaluate the clinical performance of the test and to characterize saliva based on age, gender and time from onset of symptoms. Our workflow achieved an overall sensitivity of 77.2% (n = 90), with 93.2% sensitivity, 97% specificity, and 0.895 Kappa for specimens containing >102 copies/μL (n = 77). Further analyses in saliva showed that viral load peaks in the first days of symptoms and decreases afterwards, and that viral load is ~10 times lower in females compared to males, and declines following symptom onset. NOP RT-PCR data did not yield relevant associations. This work suggests that saliva reflects the transmission dynamics better than NOP specimens, and reveals gender differences that may reflect higher transmission by males. This saliva RT-LAMP workflow can be applied to track viral spread and, to maximize detection, testing should be performed immediately after symptoms are presented, especially in females.
SUMMARYStrongyloides venezuelensis is a parasitic nematode of rats which is frequently used as a model to study human and animal strongyloidiasis. The aim of this study was to evaluate the correlation between parasitological and molecular diagnosis in Strongyloides venezuelensis infection. PCR assays were used to detect S. venezuelensis DNA in fecal samples obtained from experimentally infected Rattus norvegicus. The results showed a higher sensitivity of the PCR assay in detecting the infection compared to parasitological methods.
Strongyloides venezuelensis is a parasitic nematode of rodents that is frequently used to obtain heterologous antigens for immunological diagnosis of human strongyloidiasis. The aim of this study was to identify antigens from filariform larvae of S. venezuelensis for immunodiagnosis of human strongyloidiasis. Soluble and membrane fractions from filariform larvae of S. venezuelensis were obtained in phosphate saline (SS and SM) and in Tris-HCl buffer (TS and TM), and were analysed by Western blotting. Different antigenic components were recognized by IgG antibodies from the sera of strongyloidiasis patients. Highest recognition was observed for a 30-40 kDa mass range present in all antigenic fractions. The band encompassing this mass range was then excised and subjected to mass spectrometry for protein identification. Immunoreactive proteins identified in the soluble fractions corresponded to metabolic enzymes, whereas cytoskeletal proteins and galectins were more abundant in the membrane fractions. These results represent the first approach towards identification of S. venezuelensis antigens for use in immunodiagnostic assays for human strongyloidiasis.
Strongyloides venezuelensis is an intestinal nematode of rats, frequently used as a model for studying human and animal strongyloidiasis. In the present study, we evaluated parasitological, serological and molecular methods for the diagnosis of experimental S. venezuelensis in rats, Rattus norvegicus. Blood and faecal samples were collected and analysed up to 60 days post infection (pi) with adult worm recovery occurring from 5 to 45 days pi. Using an enzyme-linked immunosorbent assay (ELISA), serum levels of IgG antibodies increased up to 28 days pi, thereafter decreasing by day 60 pi. Polymerase chain reaction (PCR) assays detected S. venezuelensis DNA in faecal samples of rats from 5 to 21 days pi. The present study therefore represents the first step towards improving the diagnosis of experimental strongyloidiasis.
SUMMARYStrongyloidiasis is a potentially serious infection in immunocompromised patients.
Thus, the availability of sensitive and specific diagnostic methods is desirable,
especially in the context of immunosuppressed patients in whom the diagnosis and
treatment of strongyloidiasis is of utmost importance. In this study, serological and
molecular tools were used to diagnose Strongyloides stercoralis
infections in immunosuppressed patients. Serum and stool samples were obtained from
52 patients. Stool samples were first analyzed by Lutz, Rugai, and Agar plate culture
methods, and then by a quantitative real time polymerase chain reaction (qPCR). Serum
samples were evaluated by an enzyme-linked immunosorbent assay (ELISA) using a
soluble (AS) or a membrane fractions antigen (AM) obtained from alkaline solutions of
the filariform larvae of Strongyloides venezuelensis. Of the 52
immunosuppressed patients, three (5.8%) were positive for S.
stercoralis by parasitological methods, compared to two patients (3.8%)
and one patient (1.9%) who were detected by ELISA using the AS and the AM antigens,
respectively. S. stercoralis DNA was amplified in seven (13.5%)
stool samples by qPCR. These results suggest the utility of qPCR as an alternative
diagnostic tool for the diagnosis of S. stercoralis infection in
immunocompromised patients, considering the possible severity of this helminthiasis
in this group of patients.
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