Performance Of Serological Tests Research Articles

Immunodiagnosis of human hydatid disease: Where do we stand?

Cystic echinococcosis (CE) is a zoonotic parasitic infection caused by the larval stage of Echinococcus granulosus. Diagnosis of CE mainly relies on a combination of serological testing along with imaging approaches. A variety of serological methods, mainly based on hydatid cyst fluid, antigen B (AgB) and antigen 5, have been developed and used for immunodiagnosis of CE, yet their performances are not satisfactory. Although utilizing of recombinant or synthetic antigens, improved the performance of serological tests, it has not applicably overcome the problem of low sensitivity and cross reactivity, seen in the diagnosis of CE. Performances of immunodiagnostic tests based on AgB subunits are promising. The 8 kDa subunit of AgB is the most studied antigen in native, synthetic or recombinant form for diagnosis of CE. From the 5 subunits of AgB, antigen B8/1 and B8/2 provided the highest diagnostic sensitivity and specificity. Moreover, detecting of specific antibodies of IgG subclasses has improved the efficacy of immunodiagnostic tests. Among the IgG subclasses, both IgG2 and IgG4 are considered as good markers for diagnosis and IgG4 as a suitable marker for follow up of the patients. In this review an overview of immunodiagnostic methods, related antigens and their performances in the diagnosis of CE are given. The paper highlights pitfall and challenges in the serological diagnosis of CE. Moreover, limitation of currently available immunodiagnostic tests and the most recent development in the designing and application of serological assays for diagnosis of CE in human are addressed.

Within herd transmission and evaluation of the performance of clinical and serological diagnosis of foot-and-mouth disease in partially immune cattle herds

The control of foot-and-mouth disease (FMD) in vaccinated populations relies upon surveillance activities such as clinical inspections (CI) and serological monitoring. New evidence to refine current surveillance guidelines has been provided by evaluating (1) the diagnostic performance of CI and serological tests for detection of FMD virus (FMDV) non-structural proteins (NSP), and (2) the within-herd transmission of the virus in partially immune cattle. Data came from 23 affected herds during an epidemic of FMDV type O in Bolivia, in 2007. All cattle (n=957) in these herds were clinically inspected and serum samples were collected one month after the last animal with clinical signs was detected. Samples were tested for the presence of antibodies against NSP using the PANAFTOSA 3ABC-ELISA test and a subset of samples were tested using the enzyme-linked immunoelectrotransfer blot assay (EITB). Data from clinical and serological diagnoses were analysed using a Bayesian model. The sensitivity Se and specificity Sp of the tests, as well as the prevalence and the within-herd reproduction ratio R of FMDV were estimated. In addition, risk factors for infection were identified. The Se of CI, the 3ABC-ELISA and the EITB tests were estimated to be 0.30, 0.88 and 0.96 respectively. The estimated Sp, in the same order, were 0.88, 0.93 and 0.97. The within-herd prevalence of infected animals ranged from 0.04 to 0.91 and R ranged from 1.02 to 2.68. It was observed that cattle coming from areas with high vaccination coverage had a lower risk of becoming infected than home-bred cattle from the affected herds, where vaccination coverage was thought to be low. Although these estimates come from herds kept under specific conditions, they provide a reference for future surveillance design and can inform simulation models for surveillance and control of FMD in similar cattle populations.

Molecular Testing for Toxoplasma Diagnosis in Aborted Fetuses- Taleghani Maternity Hospital- Arak- Iran

Background: The diagnosis of toxoplasmosis is most critical in pregnant women who acquire infection during gestation and also in fetuses and newborns who are congenitally infected. This study described the performance of molecular and confirmatory serologic testing for toxoplasma infection in the tissues of human spontaneous aborted fetuses and their mothers' blood. Materials and Methods: 87 random samples from the tissues of body of spontaneous aborted fetuses (less than 14 weeks) in a separate container of preservative solution were collected from the delivery room of the university maternity hospital, Arak- Iran, during autumn 2012 to 2013. In the ward, 3 ml of blood sample of their mothers were collected and the sera were separated and analyzed by ELISA method for the detection of specific IgG. DNA extraction from the tissues of fetuses was performed and stored until use. The PCR reaction was performed by a pair of primers. PCR products were analyzed by electrophoresis and stained with safe stain. It is necessary to mention first that the written consent was obtained from their mothers and after recovery, a demographic questionnaire was completed. Results: Most of the mothers were 20-29 years of age and the correlation between the location of residence, contact with cats and eating undercooked food with abortion, not significant. Serological tests on the sera of 87 mothers for antiToxoplasma IgG showed 39.08% positive results. The results of PCR amplification showed that none of the 87 samples from aborted fetuses were infected with Toxoplasma gondii. Conclusion: In aborted fetuses, we did not observe any evidence of Toxoplasmosis and it appears that Toxoplasma gondii was not the cause of spontaneous abortion in this area of Iran but considering the importance of the infection during pregnancy, the control measurements during pregnancy is required.

Evaluation of a rapid serological test for leprosy classification using human serum albumin as the antigen carrier

The presence of anti-BSA antibodies may interfere in serological tests, as ELISA or immunochromatographic assays. BSA is frequently used as a blocking agent or as “inert” carrier of antigens, such as the NT-P-BSA, the semi-synthetic trisaccharide analogue of the PGL-I (phenolic glycolipid-I) antigen from the cell wall of the Mycobacterium leprae. PGL-I was prepared and linked to human serum albumin based in the hypothesis that replacing BSA by a human protein carrier would enhance the performance of leprosy serological tests. A total of 1162 serum samples were tested by ELISA and by the ML Flow rapid test using NT-P-BSA or NT-P-HSA antigens. When grouping leprosy patients as paucibacillary (PB) or multibacillary (MB) according to the Ridley & Jopling classification, ML Flow BSA and ML Flow HSA tests correctly allocated 70.9% and 68.6% of patients in the PB group, and 87% and 81% of patients in the MB group, respectively. Concordant results were found in 82.0% (953/1162) (kappa value=0.637; sd=0.023) of samples between ML Flow tests and 85.7% (996/1162) (kappa value=0.703; sd=0.021) between ELISA tests. ML Flow results were statistically similar and the same was true for ELISA tests using HSA or BSA. However, we noticed a tendency to decreased capacity to detect MB patients and an increased positivity among PB patients, HHC, TB patients and healthy controls by the HSA carrier in both ML Flow and ELISA. The PGL-I serology performed by the ML Flow test with BSA or HSA as antigen carriers can be a useful, friendly auxiliary tool to identify patients with higher bacterial load.

Issues associated with the emergence of coeliac disease in the Asia–Pacific region: a working party report of the World Gastroenterology Organization and the Asian Pacific Association of Gastroenterology.

Once thought to be uncommon in Asia, coeliac disease (CD) is now being increasingly recognized in Asia–Pacific region. In many Asian nations, CD is still considered to be either nonexistent or very rare. In recognition of such heterogeneity of knowledge and awareness, the World Gastroenterology Organization and the Asian Pacific Association of Gastroenterology commissioned a working party to address the key issues in emergence of CD in Asia. A working group consisting of members from Asia–Pacific region, Europe, North America, and South America reviewed relevant existing literature with focus on those issues specific to Asia–Pacific region both in terms of what exists and what needs to be done. The working group identified the gaps in epidemiology, diagnosis, and management of CD in Asian–Pacific region and recommended the following: to establish prevalence of CD across region, increase in awareness about CD among physicians and patients, and recognition of atypical manifestations of CD. The challenges such as variability in performance of serological tests, lack of population-specific cut-offs values for a positive test, need for expert dietitians for proper counseling and supervision of patients, need for gluten-free infrastructure in food supply and creation of patient advocacy organizations were also emphasized. Although absolute number of patients with CD at present is not very large, this number is expected to increase over the next few years or decades. It is thus appropriate that medical community across the Asia–Pacific region define extent of problem and get prepared to handle impending epidemic of CD.

Evaluation of the performance of cellular and serological diagnostic tests for the diagnosis of tuberculosis in an alpaca (Vicugna pacos) herd naturally infected with Mycobacterium bovis

Tuberculosis (TB) in llamas and alpacas has gained importance in recent years since they are imported into the European Union mainly for serving as pets and for production of natural fibre. The intradermal tuberculin test has been widely used for diagnosis of TB in these species showing lack of sensitivity (Se) although little information has been previously reported evaluating the effect on its performance of different PPD inoculation sites and time of readings. Moreover, different cost-effective serological assays have been developed in the recent years for TB diagnosis in camelids obtaining a variety of results and, for this reason, new assays still being developed. The main objectives of this study were: (1) to evaluate the performance of the intradermal tuberculin test using different inoculation sites (axillary, prescapular and cervical) and times of reading (72 and 120h) and (2) to test a novel serological assay based on MPB83 antigen in a Mycobacterium bovis naturally infected alpaca herd in Spain. In regards to skin test, single intradermal tuberculin (SIT) test at the prescapular site and reading at 72h showed the highest proportion of test-positive-culture positive animals among all culture positive animals (T+/C+), ranging from 53.8% (95% CI, 37.2–69.9) to 80% (95% CI, 44.4–97.5) using a more stringent interpretation than typically prescribed although, in general, low T+/C+ was achieved using both SIT and single comparative intradermal tuberculin (SCIT) tests alone. T+/C+ of the serological assay increased using samples collected 15–30 days after PPD injection [76.9% (95% CI, 60.7–88.9) – 100% (95% CI, 69.2–100)]. The best results of T+/C+ were obtained applying in parallel the most sensitive SIT test and serology using samples collected 15–30 days after PPD inoculation [90% (95% CI, 55.5–99.7)-100% (95% CI, 69.2–100)]. Therefore implementation of serology in parallel with the most sensitive skin test could maximize the detection of infected animals.

Melioidosis in a European traveler without comorbidities: a case report and literature review

Melioidosis is an endemic disease in Southeast Asia and northern Australia. It habitually affects immune-depressed hosts and may have a wide range of clinical manifestations. The use of positron emission tomography-computed tomography (PET-CT) has not been described previously for this disease. We report the case of a European traveler without comorbidities who developed melioidosis with pulmonary and bone marrow involvement 1 year after exposure. Antibiotic treatment was managed by taking into account the evolution on PET-CT. We review the literature and suggest the use of PET-CT for the initial evaluation of melioidosis, especially to look for a bone location, and to manage the length of antibiotic therapy.

Laboratory capacity for diagnosis of foot-and-mouth disease in Eastern Africa: implications for the progressive control pathway.

BackgroundAccurate diagnosis is pertinent to any disease control programme. If Eastern Africa is to work towards control of foot-and-mouth disease (FMD) using the Progressive Control Pathway for FMD (PCP-FMD) as a tool, then the capacity of national reference laboratories (NRLs) mandated to diagnose FMD should match this task. This study assessed the laboratory capacity of 14 NRLs of the Eastern Africa Region Laboratory Network member countries using a semi-structured questionnaire and retrospective data from the World Reference Laboratory for FMD annual reports and Genbank® through National Centre for Biotechnology Information for the period 2006–2010.ResultsThe questionnaire response rate was 13/14 (93%). Twelve out of the 13 countries/regions had experienced at least one outbreak in the relevant five year period. Only two countries (Ethiopia and Kenya) had laboratories at biosecurity level 3 and only three (Ethiopia, Kenya and Sudan) had identified FMD virus serotypes for all reported outbreaks. Based on their own country/region assessment, 12/13 of these countries /regions were below stage 3 of the PCP-FMD. Quarantine (77%) and vaccination (54%) were the major FMD control strategies employed. The majority (12/13) of the NRLs used serological techniques to diagnose FMD, seven used antigen ELISA and three of these (25%) also used molecular techniques which were the tests most frequently requested from collaborating laboratories by the majority (69%) of the NRLs. Only 4/13 (31%) participated in proficiency testing for FMD. Four (31%) laboratories had no quality management systems (QMS) in place and where QMS existed it was still deficient, thus, none of the laboratories had achieved accreditation for FMD diagnosis.ConclusionsThis study indicates that FMD diagnostic capacity in Eastern Africa is still inadequate and largely depends on antigen and antibody ELISAs techniques undertaken by the NRLs. Hence, for the region to progress on the PCP-FMD, there is need to: implement regional control measures, improve the serological diagnostic test performance and laboratory capacity of the NRLs (including training of personnel as well as upgrading of equipment and methods, especially strengthening the molecular diagnostic capacity), and to establish a regional reference laboratory to enforce QMS and characterization of FMD virus containing samples.

Poor Performance of Serological Tests in the Diagnosis of Pulmonary Tuberculosis: Evidence from a Contact Tracing Field Study

BackgroundDelayed or missed diagnosis of TB continues to fuel the global TB epidemic, especially in resource limited settings. Use of serology for the diagnosis of tuberculosis, commonly used in India, is another factor. In the present study a commercially available serodiagnostic assay was assessed for its diagnostic value in combination with smear, culture and clinical manifestations.Methodology/Principal FindingsA total of 2300 subjects were recruited for the study, but 1041 subjects were excluded for various reasons. Thus 1259 subjects were included in the study of which 470 were pulmonary tuberculosis cases (440 of 470 were culture-positive) and 789 were their asymptomatic contacts. A house-to-house survey method was used. Blood samples were tested for IgM, IgA, and IgG antibodies using the Pathozyme Myco M (IgM), Myco A (IgA) and Myco G (IgG) enzyme immunoassay (EIA). Out of 470 PTB cases, BCG scar was positive in 82.34%. The Mantoux test and smear positivity rates in PTB cases were 94.3% (430/456), and 65.32% (307/470), respectively. Among the asymptomatic contacts, BCG scar was positive in 95.3% and Mantoux test was positive in 80.66% (442/548) contacts. No contact was found falsely smear positive. The sensitivity of IgM, IgA, and IgG EIA tests was 48.7%, 25.7% and 24.4%, respectively, while the specificity was 71.5%, 80.5%, 76.6%, respectively. Performance of EIAs was not affected by the previous BCG vaccination. However, prior BCG vaccination was statistically significantly (p = 0.005) associated with Mantoux test positivity in PTB cases but not in contacts (p = 0.127). The agreement between serology and Mantoux test was not significant.ConclusionThe commercial serological test evaluated showed poor sensitivity and specificity and suggests no utility for detection of pulmonary tuberculosis.

Optimal Design of Intervention Studies to Prevent Influenza in Healthy Cohorts

BackgroundInfluenza cohort studies, in which participants are monitored for infection over an epidemic period, are invaluable in assessing the effectiveness of control measures such as vaccination, antiviral prophylaxis and non-pharmaceutical interventions (NPIs). Influenza infections and illnesses can be identified through a number of approaches with different costs and logistical requirements.Methodology and Principal FindingsIn the context of a randomized controlled trial of an NPI with a constrained budget, we used a simulation approach to examine which approaches to measuring outcomes could provide greater statistical power to identify an effective intervention against confirmed influenza. We found that for a short epidemic season, the optimal design was to collect respiratory specimens at biweekly intervals, as well as following report of acute respiratory illness (ARI), for virologic testing by reverse transcription polymerase chain reaction (RT-PCR). Collection of respiratory specimens only from individuals reporting ARI was also an efficient design particularly for studies in settings with longer periods of influenza activity. Collection of specimens only from individuals reporting a febrile ARI was less efficient. Collection and testing of sera before and after influenza activity appeared to be inferior to collection of respiratory specimens for RT-PCR confirmation of acute infections. The performance of RT-PCR was robust to uncertainty in the costs and diagnostic performance of RT-PCR and serological tests.Conclusions and SignificanceOur results suggest that unless the sensitivity or specificity of serology can be increased RT-PCR will remain as the preferable outcome measure in NPI studies. Routine collection of specimens for RT-PCR testing even when study participants do not report acute respiratory illness appears to be the most cost efficient design under most scenarios.

Assessment of performance of selected serological tests for diagnosing brucellosis in pigs

Swine brucellosis due to Brucella suis is considered an emerging zoonotic disease whose control is based on serological testing and the subsequent culling of seropositive animals or the full depopulation of affected flocks. Here we assessed the performance of several serological tests (Rose Bengal Test [RBT], indirect ELISA [i-ELISA], blocking ELISA [b-ELISA], and two competitive ELISAs [c-ELISA]) for diagnosing swine brucellosis caused by B. suis biovar 2. Both frequentistic and Bayesian statistical inference were used. A frequentistic analysis, using sera from known gold standard (GS) populations (i.e., from truly infected or brucellosis free animals), resulted in maximum (100%) diagnostic sensitivity (Se) and specificity (Sp) in the RBT, i-ELISA and b-ELISA tests. However, c-ELISAs resulted in lower diagnostic Se (ranging from 68.5% to 92.6%, according to the different cut-offs selected). A Bayesian analysis of tests yielding the best diagnostic performance with GS sera (RBT, i-ELISA and b-ELISA), but using a large collection of field sera, resulted in similar Se among tests but markedly lower (≈80%) than that resulting from the frequentistic analysis using the GS serum populations. By contrast, the estimated Sp in the Bayesian analysis was only slightly lower than 100%, thus similar to that obtained frequentistically. Our results show that adequate diagnostic tests for brucellosis in swine are available, but also emphasize the need for more extensive validation studies before applying these tests under field conditions.

Developing a new clinical tool for diagnosing chronic Q fever: theCoxiellaELISPOT: Table 1

Definitively establishing a clinical diagnosis of chronic Q fever remains challenging, as the diagnostic performance of both conventional serological tests and PCR is limited. Given the importance of an early diagnosis of chronic Q fever, there is a need for a reliable diagnostic test. We developed an enzyme-linked immunospot assay to measure Coxiella burnetii (C.burnetii)-specific T-cell responses (Coxiella ELISPOT) to both phase I and phase II antigens and tested convalescent Q fever patients (without chronic disease, n=9) and patients with an established diagnosis of chronic Q fever (n=3). The Coxiella ELISPOT adequately identified convalescent Q fever patients from healthy controls by demonstrating C.burnetii-specific T-cell interferon-γ production to both phase I and phase II antigens. Compared to convalescent Q fever patients, chronic Q fever patients showed a distinct Coxiella ELISPOT profile characterized by a much higher spot count for both phase I and phase II (18-fold for phase II, 8-fold higher for phase I) and a consistent shift towards more phase I reactivity. The diagnostic potential of the Coxiella ELISPOT is promising and warrants further investigation.

Mycoplasma pneumoniae: IgG and IgM antibody response in presence of different antigens. Evaluation of commercial tests

Background. P1 adhesin protein (170-kDa) is responsible for the interaction between M. pneumoniae and the host. Such protein is the target of specific antibodies produced by the host in response to M. pneumoniae infection. Objectives. The aim of this study was to evaluate the performance of serological tests for Mycoplasma pneumoniae IgG and IgM on chemiluminescence analyzer LIAISON® and to demonstrate how the determination of specific antibodies is the main tool for accurate diagnosis of Mycoplasma infection. Study Design. Eighty-one unselected samples were assayed by ELISA kits Vircell IgG and IgM (bacterial lysate) and LIAISON® Mycoplasma IgG and IgM (recombinant protein P1). The discordant samples were tested with ELISA Savyon Serorecombinant IgG and IgM (recombinant protein P1) and reclassified according to the consensus rule. Results. Fifty-one samples were concordant between ELISA Vircell IgG and LIAISON® Mycoplasma IgG. After resolution of the 30 discordant: twenty-eight samples were classified concordant negative and one positive. Seventy-seven samples were concordant between ELISA Vircell IgM and LIAISON® Mycoplasma IgM. After resolution of 4 discordant samples: three samples were classified as negative. Conclusions. The higher correlation between LIAISON® and Savyon Serorecombinant is determined by the use of recombinant protein P1 in the solid phase and the type of cut-off calculation allows high specificity and sensitivity. The Mycoplasma pneumoniae IgG and IgM kits, performed in full automation on LIAISON® analyzer, show characteristics comparable to other kits and an easy handling of the sample.

O2-S2.03 Feasibility of giving HSV serological test results by mail and phone

BackgroundAccurate serological tests for the herpes simplex virus have been commercially available since 1999. Despite this availability, they are offered infrequently. Concerns about the resources required for testing including time...

The Effect of Angiogenesis-related Cytokine Profiles on Rotator Cuff Disease: Discovery of New Biomarkers of Early Tendon Degeneration (SS-19)

Introduction Approximately 16% of the general population is believed to have rotator cuff disease (RCD) at a given time. The hallmarks of RCD are pro- or anti-inflammation, abnormal immune response, angiogenesis, and variables of vascularity. The relationship between angiogenesis and early degenerative changes in RCD is attracting increasing attention in orthopaedics today. Avascular zones of tendons are predisposed for degenerative changes and spontaneous rupture in RCD. Degenerative changes enhance production of pro- or anti-inflammatory and vascular angiogenesis-related cytokines (ARC) that play a pivotal role in the immune response to arthroscopic surgery and take part in the pathogenesis of RCD. Abnormalities in ARC network of importance for the clinical signs and symptoms will be focused on. To evaluate the diagnostic performance of serological tests for early degenerative changes in RCD patients: the alterations in endogenous ARC expression (IL-1beta, IL-6, IL-8, IL-10, VEGF, bFGF, ANG) in the blood. Methods The blood specimens were obtained from 200 patients with RCD from Department of Sports Medicine and Arthroscopy. Patients were followed with clinical measures (UCLA, Constant, and WORC). The comparative group consisted of 200 patients seen in the orthopedic clinic for non-tendon related shoulder complaints. The control group consisted of 200 age- and sex-matched healthy controls. Angiogenesis imaging assays was performed using PDUS for evaluation the variables of vascularity of the rotator cuff tendons. Expression ARC were measured by commercial Immunoassay kits Bio-Plex Human Cytokine Assay (Bio-Rad Inc., Hercules, CA, USA) per the manufacturer′s instructions, International Standards of the National Institute for Biological Standards and Control and recommendations of the Expert Committee on Biological Standardization of the WHO for of cytokine measurements. Affinity chromatography yielded ARC (revealed upon Ion-exchange Chromatography on QAE Sephadex) from both control and RCD patients. Results At time of inclusion the concentration of IL-1beta, IL-8, and VEGF was significantly higher in RCD patients than in control. Significantly higher serum VEGF levels were found in 85% RCD patients. VEGF overexpression correlated with advanced stage (r=0.75; p Conclusion This investigation provides the first standardized analytic approach for assessing differences in serum ARC levels of RCD. Our data demonstrated that uncontrolled pathogenesis of RCD is associated with an imbalance between inflammatory, anti-inflammatory and vascular ARC. In RCD patients was a positive correlation between ARC expression and tendon degeneration. The future of ARC in RCD application requires close interactions between the public and private sectors: the government, the pharmaceutical, biotechnology industries, and academia.

Investigation of Genotypes of Borrelia burgdorferi in Ixodes scapularis Ticks Collected during Surveillance in Canada

The genetic diversity of Borrelia burgdorferi sensu stricto, the agent of Lyme disease in North America, has consequences for the performance of serological diagnostic tests and disease severity. To investigate B. burgdorferi diversity in Canada, where Lyme disease is emerging, bacterial DNA in 309 infected adult Ixodes scapularis ticks collected in surveillance was characterized by multilocus sequence typing (MLST) and analysis of outer surface protein C gene (ospC) alleles. Six ticks carried Borrelia miyamotoi, and one tick carried the novel species Borrelia kurtenbachii. 142 ticks carried B. burgdorferi sequence types (STs) previously described from the United States. Fifty-eight ticks carried B. burgdorferi of 1 of 19 novel or undescribed STs, which were single-, double-, or triple-locus variants of STs first described in the United States. Clonal complexes with founder STs from the United States were identified. Seventeen ospC alleles were identified in 309 B. burgdorferi-infected ticks. Positive and negative associations in the occurrence of different alleles in the same tick supported a hypothesis of multiple-niche polymorphism for B. burgdorferi in North America. Geographic analysis of STs and ospC alleles were consistent with south-to-north dispersion of infected ticks from U.S. sources on migratory birds. These observations suggest that the genetic diversity of B. burgdorferi in eastern and central Canada corresponds to that in the United States, but there was evidence for founder events skewing the diversity in emerging tick populations. Further studies are needed to investigate the significance of these observations for the performance of diagnostic tests and clinical presentation of Lyme disease in Canada.

Performance of FDA-Approved Serologic Testing for Latex Allergy in an At-Risk Population

Performance of FDA-Approved Serologic Testing for Latex Allergy in an At-Risk Population

Diagnosis ofin uteroParvovirus B19 infection and maternal immune response – the relevance of linear epitopes and advanced serologic testing

Parvovirus B19 (B19V) infection in utero causing fetal anemia and non-immune hydrops fetalis (NIHF) is a potentially life-threatening event for the fetus. Postexpositional non-invasive diagnosis is based on maternal IgG/IgM response and detection of viral genome in maternal blood. Serologic testing directs prenatal follow-up. Fetal infection is confirmed by polymerase chain reaction or in situ hybridization in fetal blood and/or amniotic fluid cells. The performance of serologic tests is significant in order to direct pre- and perinatal care at rational use of resources. Timing of diagnostic procedures and knowledge of the time course of infection in pregnant, asymptomatic women are critical. IgM negative testing in the presence of prolonged viremia may complicate individual risk analysis in pregnancy. Recently, advanced IgG avidity assays and epitope-type specific assays (IgG ETS EIA) have been re-evaluated. Epidemiology, clinical relevance and management of B19V infection in pregnancy. A review of the current literature (November 1984 - May 2009) and evaluation of current information on performance and predictive value of molecular and VP1/VP2 antigen-based IgG tests directed at the diagnosis of materno-fetal B19V infection and detection of past immunity. New aspects of B19V-associated fetal disease other than anemia/NIHF are also covered. An overview of immunology and clinical relevance of B19V infection in pregnancy, of the potential value of advanced serologic testing and fields of future research. In the absence of a commercially available vaccine, serologic tests remain important tools in individual risk analysis of pregnant women exposed to B19V. Sequential application of IgG avidity and IgG ETS EIAs may improve risk stratification and timing of invasive testing in B19V-exposed pregnancies, in particular with IgM-negativity and/or persistent DNAemia. Prospective evaluation of these test systems correlated to fetal outcome in order to reduce fetal morbidity and mortality as well as the overall burden of disease of B19V with regard to fetal malformation may be subject to future research.

Celiac disease serology in patients with different pretest probabilities: Is biopsy avoidable?

To establish the diagnostic performance of several serological tests, individually and in combination, for diagnosing celiac disease (CD) in patients with different pretest probabilities, and to explore potential serological algorithms to reduce the necessity for biopsy. We prospectively performed duodenal biopsy and serology in 679 adults who had either high risk (n = 161) or low risk (n = 518) for CD. Blood samples were tested using six assays (enzyme-linked immunosorbent assay) that detected antibodies to tissue transglutaminase (tTG) and deamidated gliadin peptide (DGP). CD prevalence was 39.1% in the high-risk population and 3.3% in the low-risk group. In high-risk patients, all individual assays had a high diagnostic efficacy [area under receiving operator characteristic curves (AU ROC): 0.968 to 0.999]. In contrast, assays had a lower diagnostic efficacy (AU ROC: 0.835 to 0.972) in the low-risk group. Using assay combinations, it would be possible to reach or rule out diagnosis of CD without biopsy in 92% of cases in both pretest populations. We observed that the new DGP/tTG Screen assay resulted in a surplus compared to more conventional assays in any clinical situation. The DGP/tTG Screen assay could be considered as the best initial test for CD. Combinations of two tests, including a DGP/tTG Screen, might be able to diagnose CD accurately in different clinical scenarios making biopsy avoidable in a high proportion of subjects.

Performance of HSV-2 type specific serological tests in men in Kenya

This study compared five serological tests with Western blot from University of Washington to determine the most accurate method for detecting antibodies to herpes simplex virus type 2 (HSV-2) in a male population in Kisumu, Kenya. A random sample of 250 fishermen from 18 beaches along Lake Victoria underwent serological testing by two generations of the HerpeSelect HSV-2 ELISA (“Focus Gen 1” and “Focus Gen 2”), Kalon HSV-2 ELISA (“Kalon”), Biokit HSV-2 Rapid Test (“Biokit”), and HerpeSelect Express Rapid HSV-2 (“Express”) against the Western blot test (“WB”) as the “gold standard”. Sensitivity and specificity of tests in this population with a high prevalence of HSV-2 (58% by WB) were: Focus Gen 1: 98.6% and 63.5%; Focus Gen 2: 99.3% and 52.3%; Biokit: 66% and 90.9%; Express: 99.3% and 44.3% and Kalon: 98.6% and 85.7%. Increasing the positive cut-off value improved the specificity of the Focus Gen 2–84.9% and Kalon to 92.2%. Focus Gen 2 offered no improvement in specificity over that of Focus Gen 1. Neither rapid assay could be recommended as either a stand-alone assay or as a confirmatory test. The results of Kalon using a cut-off of 1.5 were the most concordant with those of WB for all the approaches tested. However, low positive Kalon test results should be interpreted with caution as they could reflect early seroconversion or false positive results.