BackgroundOver the last two decades, many epidemiological studies were performed to describe risks and clinical presentations of melioidosis in endemic countries.MethodsWe performed a retrospective analysis of 158 confirmed cases of melioidosis collected from medical records from 2001 to 2015 in Hospital Universiti Sains Malaysia, Kubang Kerian, Kelantan, Malaysia, in order to update the current status of melioidosis clinical epidemiology in this putatively high risk region of the country.ResultsPrincipal presentations in patients were lung infection in 65 (41.1 %), skin infection in 44 (27.8 %), septic arthritis/osteomyelitis in 20 (12.7 %) and liver infection in 19 (12.0 %). Bacteremic melioidosis was seen in most of patients (n = 121, 76.6 %). Focal melioidosis was seen in 124 (78.5 %) of patients and multi-focal melioidosis was reported in 45 (28.5 %) cases. Melioidosis with no evident focus was in 34 (21.5 %) patients. Fifty-four (34.2 %) patients developed septic shock. Internal organ abscesses and secondary foci in lungs and/or soft tissue were common. A total of 67 (41 %) cases presented during the monsoonal wet season. Death due to melioidosis was reported in 52 (32.9 %) patients, while relapses were occurred in 11 (7.0 %). Twelve fatal melioidosis cases seen in this study were directly attributed to the absence of prompt acute-phase treatment. Predisposing risk factors were reported in most of patients (n = 133, 84.2 %) and included diabetes (74.7 %), immune disturbances (9.5 %), cancer (4.4 %) and chronic kidney disease (11.4 %). On multivariate analysis, the only independent predictors of mortality were the presence of at least one co-morbid factor (OR 3.0; 95 % CI 1.1–8.4), the happening of septic shock (OR 16.5; 95 % CI 6.1–44.9) and age > 40 years (OR 6.47; 95 % CI 1.7–23.8).ConclusionsMelioidosis should be recognized as an opportunistic nonfatal infection for healthy person. Prompt early diagnosis and appropriate antibiotics administration and critical care help in improved management and minimizing risks for death.

my (M.A.H.A.); irekeola@student.usm.my (A.A.I.); hanimkk@usm.my (R.H.S.)Abstract: Epstein-Barr virus (EBV) is the causative agent of many diseases including infectious mononucleosis (IM), and it is associated with different subtypes of lymphoma, sarcoma and carcinoma such as Hodgkin's lymphoma, non-Hodgkin's lymphoma, nasopharyngeal carcinoma, and gastric carcinoma. With the advent of improved laboratory tests for EBV, a timelier and accurate diagnosis could be made to aid better prognosis and effective treatment. For histopathological lesions, the in situ hybridization (ISH) of EBV-encoded RNA (EBER) in biopsy tissues remains the gold standard for detecting EBV. Methods such as the heterophile antibody test, immunofluorescence assays, enzyme immunoassays, Western blot, and polymerase chain reaction (PCR) are also employed in the detection of EBV in different types of samples. The determination of EBV viral load using PCR, however, is gaining more prominence in the diagnosis of EBV-associated diseases. Given the challenge of false positive/negative results that are sometimes experienced during the detection of EBV, variability in results from different laboratories, and the impact of factors such as sample type and the immunological status of patients from whom samples are collected, the need to critically examine these present methods is invaluable. This review thus presents current advances in the detection of EBV, detailing the advantages and disadvantages of the various techniques. In addition, fundamental virological concepts are highlighted to enhance the greater understanding, the proper application, and the interpretation of EBV tests.Pathogens 2020, 9, 226 2 of 17 EBV can infect a wide range of cells and tissues including T and B lymphocytes, nasopharynx and oropharynx squamous epithelial cells, salivary and stomach glands, thyroid glandular epithelial cells, smooth muscle, and follicular dendritic cells [4]. However, EBV primarily infects and replicates in the stratified squamous epithelium of the oropharynx, followed by a latent infection of B lymphocytes [4]. It has been suggested that the EBV infection of B lymphocytes occurs in the oropharyngeal lymphoid organs [2]. In normal carriers, the virus persists in circulating memory B cells and initiates the production of immunoglobulins [1,2]. Following EBV's infection of B cells, a specific set of latency-related genes and transcripts are expressed, and the virus could remain dormant in resting memory B cells, from which it intermittently reactivates at any mucosal site where B cells are present (Table 1) [4,5]. The reactivation of EBV poses a great and difficult challenge to infected hosts [3]. In healthy adults, it is estimated that for every million B cells in circulation, approximately 1 to 50 are infected with EBV, with the number of latently-infected cells in each individual remaining stable for several years [6]. Therefore, EBV coexists with most human hosts without obvious outcomes. However, in some people, the virus is associated with the develo...

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Background This study reports the analytical sensitivity and specificity of a Loop-mediated isothermal amplification (LAMP) and compares its amplification performance with conventional PCR, nested PCR (nPCR) and real-time PCR (qPCR). All the assays demonstrated in this study were developed based on Serine-rich Entamoeba histolytica protein (SREHP) gene as study model. Results A set of SREHP gene specific LAMP primers were designed for the specific detection of Entamoeba histolytica. This set of primers recorded 100% specificity when it was evaluated against 3 medically important Entamoeba species and 75 other pathogenic microorganisms. These primers were later modified for conventional PCR, nPCR and qPCR applications. Besides, 3 different post-LAMP analyses including agarose gel electrophoresis, nucleic acid lateral flow immunoassay and calcein-manganese dye techniques were used to compare their limit of detection (LoD). One E. histolytica trophozoite was recorded as the LoD for all the 3 post-LAMP analysis methods when tested with E. histolytica DNA extracted from spiked stool samples. In contrast, none of the PCR method outperformed LAMP as both qPCR and nPCR recorded LoD of 100 trophozoites while the LoD of conventional PCR was 1000 trophozoites. Conclusions The analytical sensitivity comparison among the conventional PCR, nPCR, qPCR and LAMP reveals that the LAMP outperformed the others in terms of LoD and amplification time. Hence, LAMP is a relevant alternative DNA-based amplification platform for sensitive and specific detection of pathogens.