Detection Of Immunoglobulin M Antibodies Research Articles

Performance of eight commercial immunoassays for the detection of cytomegalovirus-specific IgM antibodies in pregnancy - no test fits all needs.

Detection of cytomegalovirus (CMV)-specific immunoglobulin M (IgM) antibodies as first-line serologic diagnosis plays an important role in identifying CMV primary infection during pregnancy. The performance characteristics of eight commercially available CMV IgM assays were compared. Sensitivity and IgM antibody kinetics were assessed using 100 acute phase and follow-up sera from 39 pregnant women with a well-defined onset of CMV primary infection. Specificity was analyzed using 50 well-characterized serum samples from pregnant women not infected or latently infected with CMV and from patients with other acute infections. Until 12 weeks after the onset of primary infection, four assays showed sensitivities of 100%, whereas the others had individual gaps to detect all primary infections in this time period. All assays showed a time-dependent decrease of IgM levels. More than 12 weeks after the onset of infection, the IgM-positive rates varied considerably between tests. The specificity was between 92% and 98% in all but one assay. The observed differences in the performance characteristics must be taken into account in CMV screening and diagnosis of primary infection during pregnancy.

Detection of Toxoplasmosis in Pregnant Women’s In Sabratha City of Using VIDS

Toxoplasmosis is usually asymptomatic, but can have severe consequences if It occurs in immunodeficient subject or fetuses. The diagnosis of toxoplasmosis during pregnancy is often based on maternal serological testing for IgM and IgG Anti- Toxoplasma antibodies. Persistence of IgM for long periods, posses' problems in distinguishing acute from chronic infection. The evaluation of specific IgG avidity enables more accurate dating, since avidity rises progressively during the course of infection. Test that measures immunoglobulin G (IgG) is used to determine if a person has been infected. If it is necessary to try to estimate the time of infection, which is of particular importance for pregnant women, a test which measures immunoglobulin M (IgM) is also used along with other tests such as an avidity test. Toxoplasma-specific IgG antibodies are detectable 1-3 weeks after infection and remain detectable for the life of the individual. Toxoplasma-specific IgM antibodies are also detectable 1-3 weeks after infection but generally decline to nil by one year after infection. The VIDAS® TOXO IgM (TXM) assay is intended for use on the instruments of the VIDAS family (VITEK® Immunodiagnostic Assay System) as an automated enzyme-linked fluorescent immunoassay (ELFA) for the presumptive qualitative detection of anti-Toxoplasma gondii IgM antibodies in human serum. Seventy-six (76) women in the first 16 weeks of pregnancy were tested for VIDAS IgM, IgG antibodies and VIDAS Toxo-IgG avidity. Low avidity antibodies were demonstrated in 2 (33.3%) of 6 sera positive with IgM assay and 4 (12.12%) of sera positive with IgG assay. Low avidity was also detected in 2 (3.27%) of 61 sera negative with IgM. The low avidity suggesting a recent infection, while high avidity in 3 (50%) of the 6 positive IgM and 24 (72.72%) of 33 positive IgG indicating that the Infection acquired in the distant past. These findings highlight the value of VIDAS IgG avidity when used in combination with the VIDAS IgM and IgG assays to provide a confirmatory evidence of an acute infection with a single serum specimen for pregnant women.

Ecological and subject-level drivers of interepidemic Rift Valley fever virus exposure in humans and livestock in Northern Kenya

Nearly a century after the first reports of Rift Valley fever (RVF) were documented in Kenya, questions on the transmission dynamics of the disease remain. Specifically, data on viral maintenance in the quiescent years between epidemics is limited. We implemented a cross-sectional study in northern Kenya to determine the seroprevalence, risk factors, and ecological predictors of RVF in humans and livestock during an interepidemic period. Six hundred seventy-six human and 1,864 livestock samples were screened for anti-RVF Immunoglobulin G (IgG). Out of the 1,864 livestock samples tested for IgG, a subset of 1,103 samples was randomly selected for additional testing to detect the presence of anti-RVFV Immunoglobulin M (IgM). The anti-RVF virus (RVFV) IgG seropositivity in livestock and humans was 21.7% and 28.4%, respectively. RVFV IgM was detected in 0.4% of the livestock samples. Participation in the slaughter of livestock and age were positively associated with RVFV exposure in humans, while age was a significant factor in livestock. We detected significant interaction between rainfall and elevation's influence on livestock seropositivity, while in humans, elevation was negatively associated with RVF virus exposure. The linear increase of human and livestock exposure with age suggests an endemic transmission cycle, further corroborated by the detection of IgM antibodies in livestock.

Trends in the Seroprevalence of Dengue in a Tertiary Care Hospital of North Karnataka, India

Introduction: Dengue virus of the Flaviviridae family is the causative agent of dengue fever. The Aedes aegypti mosquito is the main vector for its transmission. Though, the cases of dengue fever are mild and self-resolving, there can be fatal complications like Dengue Haemorrhagic Fever (DHF) and Dengue Shock Syndrome (DSS). Aim: To study the trends in the seroprevalence of dengue in serum samples of suspected cases. Materials and Methods: The present study was a cross- sectional study which was conducted from January 2017 to December 2019, at Belagavi Institute of Medical Sciences (BIMS), Belagavi, Karnataka, India. The serum samples were collected from suspected dengue fever cases and tested by Immunoglobulin M (IgM) capture Enzyme Linked Immunosorbent Assay (ELISA), to detect IgM antibody against dengue virus and NS1 capture ELISA for dengue NS1 (nonstructural protein 1) antigen using ELISA kits manufactured by National Institute of Virology (NIV), Pune. The tests were performed according to the manufacturer’s instruction. The data obtained from the study was analysed using descriptive statistics. Results: A total of 8,992 serum samples were tested over a period of three years, of which 1,340 (14.90%) were positive for dengue infection. Among which 1,048 (78.21%) were positive for anti-dengue IgM antibodies, 109 (8.13%) were positive for NS1 antigen and 183 (13.66%) were positive for both. Most affected age group was 11-20 years and male to female ratio is 1.18:1. The seasonal peak was observed in monsoon i.e. month of June (15.52%) followed by August (12.02%). Conclusion: Seroprevalence of dengue infection being critical signifies the importance of detection of both IgM antibodies and NS1 antigen for diagnosis of dengue infection. The study also identifies younger population being at higher risk and also monsoon as the most favourable season for viral transmission in this region and highlights the importance of concerted efforts towards disease control and prevention.

A retrospective analysis of serological & molecular testing data on dengue fever in Kolkata & adjacent districts during 2016-2019.

Regional Virus Research and Diagnostic Laboratory established at ICMR-National Institute of Cholera and Enteric Diseases (NICED) regularly receives samples for dengue screening and serotyping from patients of acute febrile illness (AFI) from Kolkata and adjacent districts. In this study, data over a three year period (August 2016-July 2019) was retrospectively analyzed to provide insight into the epidemiological trends of dengue fever in this region. Serological screening of dengue was performed by detection of NS1 antigen and/or immunoglobulin M (IgM) antibody. Dengue serotyping was done by conventional or real-time reverse transcriptase-PCR. The data were analyzed to describe the distribution of dengue with respect to age of patient, duration of fever on the day of blood collection and month of the year. Zip codes were used for spatial plotting. Out of the 24,474 samples received from Kolkata and its adjacent districts (Hooghly, Howrah, North and South 24 Parganas), 38.3 per cent (95% confidence interval: 37.7-38.9%) samples were screened positive for dengue. The correlation between age and dengue positivity was found to be weak. A combination of dengue NS1 antigen and dengue IgM antibody detection may be a better option for detecting dengue positivity compared to a single test. Most AFI cases were tested from August to November during the study period, with maximum dengue positivity noted during September (45.9%). The predominant serotype of 2016, dengue virus serotype 1 (DENV-1), was almost entirely replaced by DENV-2 in 2017 and 2018. Dengue continues to be an important cause of AFI in the region and round-the-year preventive measures are required for its control. Serotype switching is alarming and should be monitored routinely.

Duration of anti-treponemal immunoglobulin M seroreversion after successful syphilis treatment in HIV-positive and -negative patients.

Treponemal immunoglobulin M (IgM) antibody detection is currently among serologic tests used for syphilis diagnosis. However, the exact role of these antibodies is unclear. In this retrospective study of 326 (198 HIV positive and 128 negative) patients with early syphilis and positive IgM serology, data were analysed to investigate the time of IgM seroreversion after treatment and correlation with covariate factors. Median time of IgM seroreversion in the study population was 9 months (range 3-84, interquartile range 5-12). No statistically significant difference was observed between HIV-positive and -negative patients. At 12 months, 80.1% of the patients had a negative IgM test. At 6 months, 100% of HIV-positive patients had a fourfold decrease or greater in Venereal Disease Research Laboratory titres, but only 35.4% had a negative treponemal IgM. Secondary and early latent stage patients had a slower seroreversion of IgM (Hazard Ratio (HR) = 0.73, p = 0.064 and HR = 0.60, p = 0.023, respectively) than those with primary syphilis. A very strong association was observed of time to seroreversion of treponemal IgM with baseline VDRL titre (p < 0.001). Treponemal IgM antibody detection often cannot distinguish between active and successfully treated syphilis. Treponemal IgM may only be useful in the cases recommended in the guidelines, and in cases of untreated syphilis, it could support but not confirm the diagnosis.

Development and Evaluation of an Electrochemical Biosensor for Detection of Dengue-Specific IgM Antibody in Serum Samples.

Dengue is an arbovirus disease transmitted mainly by Aedes mosquitoes. As dengue shares similar clinical symptoms with other infectious diseases, prompt and accurate diagnosis is pivotal to clinicians’ decisions on appropriate management. Conventional diagnostic tests to detect the dengue-specific IgM antibody are limited in their performance and ease of use. To address these issues, we developed and evaluated a biosensor based on screen-printed carbon electrodes (SPCEs) for the detection of dengue-specific immunoglobulin M (IgM) antibodies. Various optimisations were performed in order to increase the sensitivity and specificity of the biosensor. For optimal and proper orientation of the paratope sites of goat anti-human IgM capture antibodies (GAHICA), various antibody techniques, including passive, covalent, protein A, protein G and streptavidin/biotin systems, were tested on the SPCEs. The assay reagents for the biosensor were also optimised prior to its evaluation. Analytical sensitivity evaluation was carried out using pooled sera, while analytical specificity evaluation was conducted on a panel of six non-dengue serum samples. Subsequently, diagnostic sensitivity and specificity evaluation were performed using 144 reference samples. Electrochemical current signals generated from H2O2 catalysed by HRP-labelled anti-dengue detection antibodies were measured using the chronoamperometric technique. With a limit of detection (LOD) of 106 serum dilution, the analytical sensitivity of the developed biosensor was 10 times higher than commercial ELISA. The analytical specificity of this dengue IgM biosensor was 100%. Similarly, the biosensor’s diagnostic performance was 100% for sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). These findings suggest that the developed biosensor has a great potential to be used to diagnose dengue after seroconversion.

Highlighted Prospects of an IgM/IgG Antibodies Test in Identifying Individuals With Asymptomatic Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Infection.

Covert severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections could be seeding new outbreaks. How to identify asymptomatic SARS-CoV-2 infections early has become a global focus. To explore the roles of immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies detection, nucleic acid tests, and computed tomography (CT) scanning to identify asymptomatic SARS-CoV-2 infection. The clinical data of 389 individuals with close contacts, including in general characteristics, SARS-CoV-2 etiology, serum-specific IgM and IgG antibody detection and CT imaging results, were systematically analyzed. The present study showed that only 89 of 389 individuals with close contacts were positive after the first nucleic acid test, while 300 individuals were still negative after 2 nucleic acid tests. Among the 300 individuals, 75 did not have pneumonia, and the other 225 individuals had pulmonary imaging changes. A total of 143 individuals were eventually diagnosed as having asymptomatic infection through IgM antibody and IgG antibody detection. The sensitivity, specificity, and false-negative rate of IgM and IgG antibody detection were approximately 97.1% (347 of 357), 95.3% (204 of 214), and 4.67% (10 of 214), respectively. It also indicated that during approximately 2 weeks, most individuals were both IgM positive and IgG positive, accounting for 68.57% (72 of 105). During approximately 3 weeks, the proportion of IgM-positive and IgG-positive individuals decreased to 8.57% (9 of 105), and the proportion of IgM-negative and IgG-positive individuals increased to 76.19% (80 of 105). There are highlighted prospects of IgM/IgG antibody detection as a preferred method in identifying the individuals with asymptomatic SARS-CoV-2 infection, especially combined with nucleic acid tests and pulmonary CT scanning.

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The diagnostic value of joint detection of serum IgM and IgG antibodies to 2019-nCoV in 2019-nCoV infection

Objective To investigate diagnostic value of immunoglobulin M (IgM) and immunoglobulin G(IgG) antibodies to 2019 Novel Coronavirus (2019-nCoV) in 2019-nCoV infection. Method This is a retrospective study. Serum samples were collected from 284 patients including outpatients and inpatients in Renmin Hospital of Wuhan University from January 20, 2020 to February 17, 2020. Among them 205 cases were 2019-nCoV infected patients, including 186 cases confirmed with nucleic acid test and 19 cases diagnosed by clinical symptoms and CT characteristics according to the New Coronavirus Pneumonia Control Protocol (5th edition) . A total of 79 subjects with other diseases but negative to 2019-nCoV infection were recruited as control group. Serum IgM and IgG antibodies to 2019-nCoV were measured with fully automated immunoassay technology for all subjects. Statistical significance between 2019-nCoV antibodies test and 2019-nCoV nucleic acid test was determined using χ2 tests. Result The sensitivity of serum IgM and IgG antibodies to 2019-nCoV were 70.24%(144/205) and 96.10%(197/205) respectively and specificity were 96.20%(76/79) and 92.41%(73/79) respectively. The positive and negative predictive values of 2019-nCoV antibodies were 95.63%(197/206) and 91.03% (71/78) respectively, and positive and negative predictive values of 2019-nCoV nucleic acid test were 100%(186/186) and 80.61%(79/98) respectively. The total coincidence rate of diagnosing 2019-nCoV infection between antibody tests and nucleic acid test for 2019-nCoV were 88.03%(250/284). Conclusion Joint detection of serum IgM and IgG antibodies to 2019-nCoV is an effective screening and diagnostic indicators for 2019-nCoV infection, and an effective complement to false negative results to nucleic acid test. Key words: Coronavirus; Pneumonia, viral; Immunoglobulin M; Immunoglobulin G; Coronavirus infections; Serologic tests

Diagnostic Accuracy of the InBios Scrub Typhus Detect™ ELISA for the Detection of IgM Antibodies in Chittagong, Bangladesh.

Here we estimated the accuracy of the InBios Scrub Typhus Detect™ immunoglobulin M (IgM) ELISA to determine the optimal optical density (OD) cut-off values for the diagnosis of scrub typhus. Patients with undifferentiated febrile illness from Chittagong, Bangladesh, provided samples for reference testing using (i) qPCR using the Orientia spp. 47-kDa htra gene, (ii) IFA ≥1:3200 on admission, (iii) immunofluorescence assay (IFA) ≥1:3200 on admission or 4-fold rise to ≥3200, and (iv) combination of PCR and IFA positivity. For sero-epidemiological purposes (ELISA vs. IFA ≥1:3200 on admission or 4-fold rise to ≥3200), the OD cut-off for admission samples was ≥1.25, resulting in a sensitivity (Sn) of 91.5 (95% confidence interval (95% CI: 96.8–82.5) and a specificity (Sp) of 92.4 (95% CI: 95.0–89.0), while for convalescent samples the OD cut-off was ≥1.50 with Sn of 66.0 (95% CI: 78.5–51.7) and Sp of 96.0 (95% CI: 98.3–92.3). Comparisons against comparator reference tests (ELISA vs. all tests including PCR) indicated the most appropriate cut-off OD to be within the range of 0.75–1.25. For admission samples, the best Sn/Sp compromise was at 1.25 OD (Sn 91.5%, Sp 92.4%) and for convalescent samples at 0.75 OD (Sn 69.8%, Sp 89.5%). A relatively high (stringent) diagnostic cut-off value provides increased diagnostic accuracy with high sensitivity and specificity in the majority of cases, while lowering the cut-off runs the risk of false positivity. This study underlines the need for regional assessment of new diagnostic tests according to the level of endemicity of the disease given the high levels of residual or cross-reacting antibodies in the general population.

Chikungunya virus infection in Aruba: Diagnosis, clinical features and predictors of post-chikungunya chronic polyarthralgia.

BackgroundChikungunya virus (CHIKV) emerged in Aruba for the first time in 2014. We studied the clinical presentation of acute CHIKV infection and the contribution of serologic and molecular assays to its diagnosis. In a cohort of confirmed CHIKV cases, we analysed the frequency, duration and predictors of post-chikungunya chronic polyarthralgia (pCHIK-CPA), defined as joint pains lasting longer than 6 weeks or longer than 1 year.MethodologyPatient sera obtained within 10 days of symptom onset were tested for CHIKV, using an indirect immunofluorescence test for the detection of CHIKV-specific Immunoglobulin M (IgM) and post-hoc, by reverse-transcription polymerase chain reaction (RT-PCR). CHIKV was isolated from selected samples and genotyped. For confirmed CHIKV cases, clinical data from chart review were complemented by a Telephone survey, conducted 18–24 months after diagnosis. When joint pain was reported, the duration, presence of inflammatory signs, type and number of joints affected, were recorded. Joint involvement was scored according to the 2010 ‘American College of Rheumatology/ European League Against Rheumatism’ criteria for seronegative rheumatoid arthritis (ACR-score). Risk factors for pCHIK-CPA were identified by logistic regression.Principal findingsAcute CHIKV infection was diagnosed in 269 of 498 sera, by detection of IgM (n = 105), by RT-PCR (n = 59), or by both methods (n = 105). Asian genotype was confirmed in 7 samples. Clinical data were complete for 171 of 248 (69.0%) patients, aged 15 years or older (median 49.4 [35.0–59.6]). The female-to-male ratio was 2.2. The main acute symptoms were arthralgia (94%), fever (85%), myalgia (85%), headache (73%) and rash (63%). In patients with arthralgia (n = 160), pCHIK-CPA longer than 6 weeks was reported by 44% and longer than 1 year by 26% of cases. Inflammatory signs, stiffness, edema and redness were frequent (71%, 39% and 21%, respectively). Joints involved were knees (66%), ankles (50%), fingers (52%), feet (46%), shoulders (36%), elbows (34%), wrists (35%), hips (31%), toes (28.1%) and spine (28.1%). Independent predictors of pCHIK-CPA longer than 1 year were female gender (OR 5.9, 95%-CI [2.1–19.6]); high ACR-score (7.4, [2.7–23.3]), and detection of CHIKV-RNA in serum beyond 7 days of symptom onset (6.4, [1.4–34.1].ConclusionsWe identified 269 CHIKV patients after the first outbreak of Asian genotype CHIKV in Aruba in 2014–2015. RT-PCR yielded 59 (28%) additional CHIKV diagnoses compared to IgM antibody detection alone. Arthralgia, fever and skin rash were the dominant acute phase symptoms. pCHIK-CPA longer than 1 year affected 26% of cases and was predicted by female gender, high ACR-score and CHIKV-RNA detection beyond 7 days of symptom onset.

Comparison of two chemiluminescent immunoassays in the detection of measles IgM antibodies

Serological confirmation of measles is achieved by detecting the specific immunoglobulin M (IgM), and it is important to evaluate new commercial inmunoassays in order to ensure the quality of results. The objective of this study was to compare the performance of a novel automated chemiluminescent immunoassay (CLIA), Virclia IgM measles (Vircell, Spain), with that of the widely used Liaison measles IgM assay (DiaSorin, Italy). A panel of 86 sera from laboratory-confirmed cases was used for the sensitivity calculation, and 59 sera from healthy individuals and those with other viral infections were used for the specificity calculation. Sensitivity values were 96.5% for Virclia and 97.6% for Liaison; specificity values were 93.2% for Virclia and 96.6% for Liaison; neither difference was statistically significant VirClia IgM measles is a good alternative to other immunoassays for the serological confirmation of measles.

Performance Characteristics of Commercial Immunoassays for the Detection of IgG and IgM Antibodies to β2 Glycoprotein I and an Initial Assessment of Newly Developed Reference Materials for Assay Calibration.

To investigate the performance characteristics and impact of newly developed reference calibrators on the commutability between anti-β2 glycoprotein I (anti-β2 GPI) immunoassays in antiphospholipid syndrome (APS) and/or systemic lupus erythematosus (SLE). Immunoglobulin G (IgG) and immunoglobulin M (IgM) anti-β2 GPI immunoassays from four manufacturers were evaluated. Serum samples from 269 patients (APS only, n = 31; SLE and APS, n = 83; SLE only, n = 129; pregnancy-related clinical manifestations without APS, n = 26) and 162 women with histories of successful pregnancies were tested. Results were expressed in kit-specific arbitrary units and in the calibrator reference units (RUs) based on 99th percentile cutoff values. Diagnostic accuracies, correlation between kits, and specific clinical manifestations in APS were investigated. The sensitivities of the assays ranged from 15.8% to 27.2% (IgG) and 12.3% to 15.8% (IgM) while specificities ranged from 79.4% to 86.5% (IgG) and 80.6% to 84.5% (IgM). There was moderate to almost perfect interassay reliability (Cohen κ, 0.69-0.98), and Spearman correlation coefficients were generally improved when results of the IgG determinations were expressed in RUs. Although qualitative agreements between immunoassays for both antibody isotypes are acceptable, correlations with APS clinical manifestations were kit dependent. Only the use of IgG reference material improved quantitative correlations between assays.

Improved Diagnosis of Acute Pulmonary Histoplasmosis by Combining Antigen and Antibody Detection.

Acute pulmonary histoplasmosis can be severe, especially following heavy inoculum exposure. Rapid diagnosis is critical and often possible by detection of antigen, but this test may be falsely negative in 17% of such cases. Antibody detection by enzyme immunoassay (EIA) may increase sensitivity and permit the measurement of immunoglobulin M (IgM) and immunoglobulin G (IgG) classes of antibodies separately. Microplates coated with Histoplasma antigen were used for testing of serum from patients with acute pulmonary histoplasmosis and controls in the MVista Histoplasma antibody EIA. Results for IgG and IgM were reported independently. IgG antibodies were detected in 87.5%, IgM antibodies in 67.5%, and IgG and/or IgM antibodies in 88.8% of patients with acute pulmonary histoplasmosis in this assay, while immunodiffusion, complement fixation, and antigen testing showed sensitivities of 55.0%, 73.1%, and 67.5%, respectively (n = 80). Combining antigen and antibody detection increased the sensitivity to 96.3%. The MVista Histoplasma antibody EIA offers increased sensitivity over current antibody tests while also allowing separate detection of IgG and IgM antibodies and complementing antigen detection. Combining antigen and EIA antibody testing provides an optimal method for diagnosis of acute pulmonary histoplasmosis.

Detection of IgM and IgG antibodies against hepatitis E virus in donated blood bags from a national voluntary blood bank in Metro Manila, Philippines

ObjectiveTo assess the blood bags obtained from a national voluntary blood bank for the presence of immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies against hepatitis E virus (HEV). MethodsPlasma from the blood bags provided by a voluntary blood bank was examined for the IgM and IgG antibodies against HEV through the ELISA technique. ResultsResults showed HEV IgG and IgM antibodies with prevalence of 11.8% and 2.4% positives, respectively. ConclusionsScreening of HEV antibodies of donors is a must to safeguard blood products and reduce the possible transmission of HEV infections.

Antibody responses of Chlamydophila pneumoniae pneumonia: Why is the diagnosis of C. pneumoniae pneumonia difficult?

The ELNAS Plate Chlamydophila pneumoniae commercial test kit for the detection of anti-C. pneumoniae-specific immunoglobulin M (IgM), IgA and IgG antibodies has become available in Japan recently. To determine the optimum serum collection point for the ELNAS plate in the diagnosis of C.pneumoniae pneumonia, we analyzed the kinetics of the antibody response in patients with laboratory-confirmed C.pneumoniae pneumonia. We enrolled five C.pneumoniae pneumonia cases and collected sera from patients for several months. The kinetics of the IgM and IgG antibody responses were similar among the five patients. Significant increases in IgM and IgG antibody titer between paired sera were observed in all patients. IgM antibodies appeared approximately 2-3 weeks after the onset of illness, reached a peak after 4-5 weeks, and were generally undetectable after 3-5 months. IgG antibodies developed slowly for the first 30 days and reached a plateau approximately 3-4 months after the onset of illness. The kinetics of IgA antibody responses were different among the five patients, and significant increases in IgA antibody titer between paired sera were observed in only two patients. Although the sample size was small, the best serum collection time seemed to be approximately 3-6 weeks after onset of illness when using a single serum sample for the detection of IgM antibodies. Paired sera samples should be obtained at least 4 weeks apart. IgA antibody analysis using ELNAS may not be a useful marker for acute C. pneumoniae pneumonia.

English

Dengue has become a major health concern globally in recent decades. Dengue infected patients manifest a spectrum of symptoms and in severe cases the fate is mortality. A hallmark of dengue infection is thrombocytopenia which causes concern for the patients and treating doctors. This study aimed to evaluate the incidence of thrombocytopenia in seropositive dengue patients. Bleeding manifestation was also investigated in dengue patients to evaluate its association with the severity of thrombocytopenia. In this study, 750 individuals were screened for dengue infection by detecting immunoglobulin M (IgM) against dengue virus in their serum. Enzyme-linked immunosorbent assay (ELISA) was performed for detection of IgM antibody and 250 individuals were found to be seropositive. Platelet counts were performed on whole blood of seropositive patients using Sysmex XE-5000 Automated Hematology Analyzer. Among 250 dengue patients, 2% had severe thrombocytopenia, 65.2% were found to have mild to moderate thrombocytopenia and 32.8% had normal platelet counts. Bleeding was related to the severity of thrombocytopenia as 80% of patients having platelet count lower than 25000/&mu;l showed bleeding manifestations. &nbsp; Key words:&nbsp; Dengue, thrombocytopenia, seropositive, immunoglobin M (IgM), enzyme-linked immunosorbent assay (ELISA), platelet count.

First Outbreak of West Nile Virus Neuroinvasive Disease in Humans, Croatia, 2012

Between September 6 and 21, 2012, seven human cases of West Nile virus (WNV) neuroinvasive infection were laboratory confirmed in Croatia. The median patient age was 62 years (range 48-77). Five patients presented with meningoencephalitis and two patients with meningoencephalitis followed by acute flaccid paralysis. Four of them had an underlying disease (hypertension). Using enzyme-linked immunosorbent assay (ELISA), WNV-specific immunoglobulin M (IgM) and IgG antibodies of low avidity were detected in six patients, whereas one showed only IgM antibodies. All samples were confirmed using plaque-reduction neutralization and microneutralization tests. Five patients recovered fully. Before human cases were reported, acute asymptomatic WNV infection was demonstrated by detection of IgM antibodies in sentinel horses. Moreover, an increased WNV IgG seropositivity in horses was detected in counties where human cases occurred. Adulticidal and larvicidal treatments were administered immediately in the respective places of residence. The end of the warm season contributed to the fact that there were no new cases of WNV disease recorded.

Antenatal screening forToxoplasma gondiiinfection at a tertiary care hospital in Riyadh, Saudi Arabia

BACKGROUND AND OBJECTIVES:Congenital toxoplasmosis is associated with significant morbidity and mortality. This study investigates the prevalence of toxoplasmosis among pregnant women.DESIGN AND SETTING:A retrospective study at King Khalid University Hospital, Riyadh from September 2009 to August 2010.PATIENTS AND METHODS:Laboratory data of 2176 pregnant women screened for Toxoplasma gondii in the antenatal care unit were assessed during the study period. The mean (SD) age of the women and the duration of pregnancy were 25 (7.3) years and 18 (7.7) weeks, respectively. Data were extracted for the presence or absence of anti-T gondii immunoglobulin G (IgG) and IgM antibodies.RESULTS:Of 2176 sera tested, 1351 (62%) did not show any evidence of exposure to T gondii. The remaining 825 (38%) samples tested positive for anti-T gondii IgG antibodies, and none was found to have anti-T gondii IgM antibodies in the serum. These data reveal that a significantly high number of women in the antenatal care unit at King Khalid University Hospital in Riyadh had been exposed to T gondii.CONCLUSION:A high prevalence of toxoplasmosis among pregnant women warrants multicenter community-based investigations for assessment of T gondii infection and identification of risk factors for transmission of toxoplasmosis in general, and particularly during pregnancy.