Evaluation and optimization of a commercial enzyme linked immunosorbent assay for detection of IgA antibodies-2

<p><b>Copyright information:</b></p><p>Taken from "Evaluation and optimization of a commercial enzyme linked immunosorbent assay for detection of IgA antibodies"</p><p>http://www.biomedcentral.com/1471-2334/8/98</p><p>BMC Infectious Diseases 2008;8():98-98.</p><p>Published online 26 Jul 2008</p><p>PMCID:PMC2515311.</p><p></p

Evaluation and optimization of a commercial enzyme linked immunosorbent assay for detection of IgA antibodies-1

Bution of SeroCP IgA index in relation to MIF IgA antibody seropositivity in the control group. The continuous lines are the manufacturer's cut off indices, and the discontinuous lines are the optimized cut off values.<p><b>Copyright information:</b></p><p>Taken from "Evaluation and optimization of a commercial enzyme linked immunosorbent assay for detection of IgA antibodies"</p><p>http://www.biomedcentral.com/1471-2334/8/98</p><p>BMC Infectious Diseases 2008;8():98-98.</p><p>Published online 26 Jul 2008</p><p>PMCID:PMC2515311.</p><p></p

Evaluation and optimization of a commercial enzyme linked immunosorbent assay for detection of IgA antibodies-0

E differences between OD 450 nm values were less than 20%.<p><b>Copyright information:</b></p><p>Taken from "Evaluation and optimization of a commercial enzyme linked immunosorbent assay for detection of IgA antibodies"</p><p>http://www.biomedcentral.com/1471-2334/8/98</p><p>BMC Infectious Diseases 2008;8():98-98.</p><p>Published online 26 Jul 2008</p><p>PMCID:PMC2515311.</p><p></p

Comparison of Two Quantitative Real Time PCR Assays for <i>Rickettsia</i> Detection in Patients from Tunisia

<div><p>Background and objectives</p><p>Quantitative real time PCR (qPCR) offers rapid diagnosis of rickettsial infections. Thus, successful treatment could be initiated to avoid unfavorable outcome. Our aim was to compare two qPCR assays for <i>Rickettsia</i> detection and to evaluate their contribution in early diagnosis of rickettsial infection in Tunisian patients.</p><p>Patients and methods</p><p>Included patients were hospitalized in different hospitals in Tunisia from 2007 to 2012. Serology was performed by microimmunofluorescence assay using <i>R. conorii</i> and <i>R. typhi</i> antigens. Two duplex qPCRs, previously reported, were performed on collected skin biopsies and whole blood samples. The first duplex amplified all <i>Rickettsia</i> species (PanRick) and <i>Rickettsia typhi</i> DNA (Rtt). The second duplex detected spotted fever group <i>Rickettsiae</i> (RC00338) and typhus group <i>Rickettsiae</i> DNA (Rp278).</p><p>Results</p><p>Diagnosis of rickettsiosis was confirmed in 82 cases (57.7%). Among 44 skin biopsies obtained from patients with confirmed diagnosis, the first duplex was positive in 24 samples (54.5%), with three patients positive by Rtt qPCR. Using the second duplex, positivity was noted in 21 samples (47.7%), with two patients positive by Rp278 qPCR. Among79 whole blood samples obtained from patients with confirmed diagnosis, panRick qPCR was positive in 5 cases (6.3%) among which two were positive by Rtt qPCR. Using the second set of qPCRs, positivity was noted in four cases (5%) with one sample positive by Rp278 qPCR. Positivity rates of the two duplex qPCRs were significantly higher among patients presenting with negative first serum than those with already detectable antibodies.</p><p>Conclusions</p><p>Using qPCR offers a rapid diagnosis. The PanRick qPCR showed a higher sensitivity. Our study showed that this qPCR could offer a prompt diagnosis at the early stage of the disease. However, its implementation in routine needs cost/effectiveness evaluation.</p></div

TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labeling (TUNEL) assay of spermatozoa.

<p>Histograms show: (<b>A</b>) negative control with 2.35% TUNEL positive cells. (<b>B</b>) Positive control (spermatozoa treated with DNaseI) with 90.5% TUNEL positive cells. (<b>C</b>) Semen sample of one male partner of infertile couples positive for <i>C. trachomatis</i> qPCR with 20.5% TUNEL positive cells. B: window adjusted to detect the percentage of TUNEL positive cells.</p

Seminological variables of semen of <i>C. trachomatis</i> positive patients compared to uninfected patients.

<p>Note: Values are means (± Standard Error).</p>#<p>Unless indicated, variables were tested T-Test.</p

Flow cytometric of changes in the mitochondrial membrane potential (ΔΨm).

<p>Histograms show: (<b>A</b>) Negative control with 95.8% low ΔΨm cells. (<b>B</b>) Positive control with 9.6% low ΔΨm cells. (<b>C</b>) Semen sample of one male partner of infertile couples positive for <i>C. trachomatis</i> qPCR with 32.5% low ΔΨm cells and 67.5% with high ΔΨm cells. C: window adjusted to detect the percentage of cells with low ΔΨm.</p

Flow cytometric of sperm viability using 7-amino-actinomycin-D Dye.

<p>Histograms show: (<b>A</b>) Negative control with 10% 7-AAD positive cells. (<b>B</b>) Positive control with 98.5% 7-AAD positive cells. (<b>C</b>) Semen sample of one male partner of infertile couples positive for <i>C. trachomatis</i> qPCR with 56.5% 7-AAD negative cells and 43.5% 7-AAD positive cells. B: window adjusted to detect the percentage of cells with 7-AAD positive.</p

Positivity of qPCR according to serologic status among patients with confirmed diagnosis.

<p>Positivity of qPCR according to serologic status among patients with confirmed diagnosis.</p

Standard curves constructed for the four quantitative PCRs.

<p>Standard curves constructed for the four quantitative PCRs.</p