Problems encountered in conventional HIV 1/2 Algorithms: lack of necessity for immunoblot assays to confirm repeated ELISA reactive results

Background: The use of conventional (serologically based) HIV 1/2 diagnostic algorithms has become controversial in recent years.Objectives: Sera from patients who underwent verification tests were evaluated because repeated ELISA-reactive results demonstrated a HIV1+HIV2 positive band pattern.Methods: The line immunoassay (LIA) test was used for repeated HIV enzyme immunoassays (EIA)-reactive sera in patients at three centers. The Bio-Rad Geenius™ HIV 1/2 and the HIV-1 RNA tests were used. HIV-1 and RNA HIV-2 were investigated using PCR.Results: LIA was used to evaluate 3,224 out of 10,591 samples with repeated ELISA reactivity (30%). We found that 32 (1%) of the sera, along with HIV1 bands and HIV2 gp36 bands, were positive. Only 28 of the 32 verified serum samples with gp36 bands were repeated, and no gp36 band positivity was detected using the Bio-Rad Geenius™ HIV-1/2 confirmatory assay in these serum samples. The HIV-2 proviral DNAs were also negative. Therefore, we excluded the possibility of HIV1+2 co-infection. All samples from the 32 patients were positive for HIV-1 RNA.Conclusion: Our findings highlight the need to exclude confirmatory tests like the LIA test from the current diagnostic HIV algorithm and replace it with rapid HIV-1 and HIV-2 confirmatory immunochromotographic tests.Keywords: HIV, AIDS, HIV-2

Problems encountered in conventional HIV 1/2 Algorithms: lack of necessity for immunoblot assays to confirm repeated ELISA reactive results

Background: The use of conventional (serologically based) HIV 1/2 diagnostic algorithms has become controversial in recent years. Objectives: Sera from patients who underwent verification tests were evaluated because repeated ELISA-reactive results demonstrated a HIV1+HIV2 positive band pattern. Methods: The line immunoassay (LIA) test was used for repeated HIV enzyme immunoassays (EIA)-reactive sera in patients at three centers. The Bio-Rad Geenius\u2122 HIV 1/2 and the HIV-1 RNA tests were used. HIV-1 and RNA HIV-2 were investigated using PCR. Results: LIA was used to evaluate 3,224 out of 10,591 samples with repeated ELISA reactivity (30%). We found that 32 (1%) of the sera, along with HIV1 bands and HIV2 gp36 bands, were positive. Only 28 of the 32 verified serum samples with gp36 bands were repeated, and no gp36 band positivity was detected using the Bio-Rad Geenius\u2122 HIV-1/2 confirmatory assay in these serum samples. The HIV-2 proviral DNAs were also negative. Therefore, we excluded the possibility of HIV1+2 co-infection. All samples from the 32 patients were positive for HIV-1 RNA. Conclusion: Our findings highlight the need to exclude confirmatory tests like the LIA test from the current diagnostic HIV algorithm and replace it with rapid HIV-1 and HIV-2 confirmatory immunochromotographic tests

Essential thrombocythemia with Mpl W515 K mutation in a child presenting with Budd-Chiari syndrome

WOS: 000369892400016PubMed ID: 25970554Essential thrombocythemia (ET) is an extremely rare childhood disorder characterised by clonal expansion of megakaryocytic lineage in bone marrow, leading to a persistent increase in the number of circulating thrombocytes and thus increased risk for thrombotic and haemorrhagic events. The molecular mechanisms of ET are not fully understood. Most children with ET have the JAK2 V617F somatic mutation; however, another mutation, involving a W to L or K substitution at Mpl codon 515, was reported in a small proportion of adult ET patients that is extremely rare in children. Herein, we describe a Mpl W515K somatic mutation in a paediatric case of ET who presented with Budd-Chiari syndrome. No paediatric patient harbouring a Mpl W515K mutation has been previously reported

New approaches to in vitro diagnosis of hepatitis C infection a reason for post transfusion hepatitis: Diagnostic value of determination of hepatitis C virus core antigen

In between the dates of February 2008-March 2009, by applying to Istanbul University CTF Microbiology and Clinical Microbiology Basic Sciences Branch and Duzen laboratories, 123 cases, where HCV RNA and anti-HCV positivity are identified with molecular (real-time PCR) and serologic (ELISA) methods as a positive control group, and 48 cases where FICV RNA and anti-HCV negativity are identified as a negative control group are established. The values of sensitivity, specificity, positive and negative approximation of recently developed HCV Core Ag (Abbott Diagnostics, Germany) kit are determined successively as 94.3%, 97.9%, 99.1%, 87%, 95.3% and 88%. Although the new HCV Ag assay is clearly not sensitive enough to replace HCV NAT it may serve as a valuable tool in the HCV diagnostic algorithm as it is able to pick up a great majority of anti-HCV and HCV RNA positive samples, thus allowing a timely and less expensive serological diagnosis of an active HCV infection. This may be an advantage for labs that do not have access to PCR easily. (C) 2011 Elsevier Ltd. All rights reserved

Detection of Intestinal Parasites with Conventional and Molecular Methods in Follow-up HIV/AIDS Cases

In people living with human immunodeficiency virus (HIV), several complaints related to the gastrointestinal system, mainly diarrhea can be determined. In our study, we aimed to detect the existence of intestinal parasites with conventional methods based on microscopy and with molecular methods based on multiplex-PCR among 90 anti-retroviral treatment (ART) naive or ART adherent HIV/AIDS cases. The existence of Giardia spp., Blastocystis spp., Entamoeba histolytica, Dientamoeba spp. and Cryptosporidium spp. were searched in stool samples and the relation with the existence of these parasites and demographic/clinical data of the cases were determined. The demographic and clinical data of the participants included in the study were as follows; the average age was 34.02 +/- 9.7 years, average time of diagnosis was 2.4 +/- 1.7 years. Gender distribution was as follows; 85.6% male and 14.4% female. HIV transmission was related with heterosexual intercourse in 60%, homosexual intercourse in 33.3%, blood/blood products contact in 1.1% and with unknown routes in 5.6% of the cases. Fifty percent of the patients were in pre-ART and 50% was in on-ART state. The average CD4+ T lymphocyte count was detected as 400 cells/mm(3) and the median of viral load was 114.527 copies/ml. An overall prevalence of at least one intestinal parasitic infection was recorded as 36.7% and the prevalence of this infection due to Blastocystis spp. was 22.2%, followed by Dientamoeba spp. (13.3%), E. histo lytica (4.4%), Cryptosporidium spp. (3.3%), Giardia spp. (2.2%) and multiple parasitic infections (7.7%). The type of sexual behaviours related with the detection of intestinal parasites were statistically significant especially in homosexual intercourse (p< 0.001). The increase in CD4+ T lymphocyte counts were reversely associated (p= 0.062) and the increase in the levels of viral load were positively associated (p< 0.001) with detection rate of intestinal parasite. The detection of parasites by molecular methods was statistically significant in pre-ART participants (p= 0.002) and participants with diarrhea (p= 0.019). In the present study, the increase in the frequency of intestinal parasitic infections has shown that essential interventions are required. In all HIV/AIDS cases, routine parasitic screening should be performed by more sensitive methods to manage early and specific treatment

Microorganisms in Respiratory Tract of Patients Diagnosed with Atypical Pneumonia: Results of a Research Based on the Use of Reverse Transcription Polymerase Chain Reaction (RT-PCR) DNA Microarray Method and Enzyme-Linked Immunosorbent Assay

Background: Numerous molecular-based tests were applied for the laboratory-based diagnosis of viruses. In this cross-sectional case control study, in addition to bacteria, we aimed to determine respiratory viruses using, for the first time in our country, the Reverse Transcription PCR DNA Microarray method, and we also aimed to evaluate its diagnostic performance

Neopterin and soluble CD14 (sCD14) levels as immunoactivation markers in aHBc-only cases

Aim: Isolated hepatitis B core antibodies (aHBc)-only pattern complicates the diagnosis of HBV infections. We evaluated neopterin and sCD14 levels in HBV infections. Methods: aHBc-only (n: 102), healthy control (healthy control group [HCG], n: 100), and chronic hepatitis (CHB) groups (n: 70) were investigated. Competitive and sandwich ELISA were used. Results: The mean neopterin levels were significantly lower in the aHBc-only group than those in the CHB group (p = 0.0001). No significant difference was found between the aHBc-only group and the HCG (p = 0.854). The mean sCD14 levels were lower in the aHBc-only group than those in the CHB group (p = 0.0001), but no significant difference was found between the aHBc-only group and HCG (p = 0.402). No significant difference was detected between aHBc-only and HCG for mean sCD14 (p = 0.402) and neopterin levels (p = 0.854). Conclusion: These two biomarkers are not useful for diagnosing the aHBc only pattern