Reverse transcription PCR assays for detection of borna disease virus in psychiatric patients

Borna disease virus [BDV] is a virus that naturally infects the central nervous system [CNS] of broad range of warm-blooded animals. BDV is an enveloped virus, non- segmented, negativestranded RNA genome and has an organization characteristic of a member of Bornaviridae. It may persists in the central nervous system [CNS] of infected individuals for their entire life span. In the present work the presence of BDV p24 RNA in peripheral blood cells was detected from 37 psychiatric patients [15 patients diagnosed as schizophrenia, 10 as Major depressive disorder and 12 as mood disorder bipolar I most recent episode mania]. Patients were selected from the outpatient as well as the inpatient units of Psychiatric Department in Mansoura University hospital and 37 healthy volunteers as the control group. All subjects were interviewed by structured diagnostic criteria categorized according to the Diagnostic and Statistical Manual of Mental Disorders DSM-IV. The presence of BDV p24 RNA was investigated by nested reverse transcriptase PCR [RT-PCR] using specific primers to p24 from BDV. The median duration of illness was 6, 4 and 10 years in depressive disorder, mood disorder and schizophrenia respectively. The mean age was 40.7, 30.1 and 36.1 in depressive disorder, mood disorder and schizophrenia respectively. There were no significant differences in age and duration of illness among patients groups with psychotic disorders in the presence or absence of p24 RNA of BDV. Frequency of BDV- RNA on patient's groups was 10.8% [4/37]. The detection of BDV-RNA in the peripheral blood cells of patients but not on control group should help our understanding of the pathogenesis in the disease

Evaluation of dermatophyte identification medium for recovery of keratinophillic fungi from clinical samples

Dermatophyte test media [DTM] and Sabouraud's dextrose agar [SDA] are well established media for recovery of dermatophytes from clinical samples of dermatophytosis. Skin, hair and nail samples were mycologically examined from 112 patients attending Outpatient Clinic of Dermatology and Veneriology in Mansoura University Hospital in an attempt to compare dermatophyte identification media [DIM] with DTM and SDA for recovery and identification of dermatophytes. One hundred and sixteen fungi were recovered from 112 patients. The most common clinical lesions were onychomycosis 35.7% [40/112] and tinea capitis 17.86% [20/112]. The most common recovered keratinophilic fungi were T. mentagrophytes [31/116], followed by T. rubrum [15/116]. On comparing DIM with SDA and DTM sensitivity was 95.74% and 94.74% respectively while specificity of DIM with DTM was 86.67%.So DIM culture is an inexpensive, rapid, specific, and accurate method for the presumptive recovery of dermatophytes from clinical samples

Laboratory methods for detection of herpes simplex virus-1in herpetic keratitis patients

Herpetic ocular disease is a major cause of blindness. Rapid and accurate diagnosis is essential for prompt, proper treatment. The purpose of this study was to evaluate molecular methods as rapid diagnostic tools compared with cell culture of HSK. Corneal scrapings from [55] patients with clinically typical dendritic corneal ulcer suggestive of HSV keratitis, and [20] patients with clinically non-viral corneal ulcers as control, were tested by Giemsa stain for multinucleated giant cells, immunofluorescence assay [IFA] for HSV-1 antigen, and polymerase chain reaction [PCR] for HSV-1 DNA, and an attempt for viral isolation was performed on Vero cell line culture. Positive samples by cell culture were 27.3%, whereas PCR was positive in [34.6 %] and IFA was positive in [29%.] PCR had better sensitivity [100%] than IFA [70%]; however, IFA had better specificity [88%] and the difference was significant; P>.001. Their sensitivity were significantly higher than Giemsa stain. This indicates that a combination of IFA and PCR constitute the choice of tests in clinically suspected cases of HSV keratitis, multinucleated giant cells in Giemsa stain may give Presumptive diagnosis of viral infection

Molecular evidence of helicobacter species in human gastric biopsy specimens

The genus Helicobacter consists of 24 formally recognized species, 7 of which have been associated with human disease. The best studied of these is the gastric pathogen Helicobacter pylori. To identify different Helicobacter [H.] species as causative organisms gastritis. 35 patients undergoing upper gastrointestinal endoscopy were selected [21males and 14 females. Age ranged from [25-60 years]. Gastric biopsy specimens were subjected to rapid urease test [RUT], bacteriological identification by Gram stain, culture and molecular detection of Helicobacter by polymerase chain reaction [PCR]. Then species identification using restriction fragment length polymorphism [RFLP] technique. 14 Helicobacter were identified [12 H. pylori, 2H. non-pylori ] were positive by PCR. Gram negative spiral organisms were detected in 10 out of 14 cases that were positive by PCR [the gold standard method] with sensitivity of 71% and a specificity of 90% while culture was positive in 10/ 14 positive PCR cases with sensitivity [71%] and high specificity of [100%] [p <0.001] and 11 out of 14 are positive by RUT, and 2 out of 14 are H. non-pylori .It was concluded that PCR. RFLP are useful molecular techniques for rapid identification and differentiation of different Helicobacter species

Helicobacter pylori CagA genotype in patients with gastritis and peptic ulcer

H. pylori infection of the stomach is widespread and is considered to play a major role in the pathogenesis of gastric diseases such as peptic ulcer and adenocarcinoma. The virulence of this type of bacteria in gastroduodenal disease is related to virulence gene [cagA] present in some strains. The aim of this study was to investigate the presence of H. pylori cagA genotype and its association with clinical outcomes. H. pylori were isolated and rapid urease test was conducted from gastric biopsy specimens obtained from 93 patients [68 with gastritis and 25 with gastric ulcer]. The cagA genotype was detected by PCR. CagA genotype was present in 41% of the patients infected with H. pylori positive both by culture and rapid urease test. CagA genotype was correlated with the severity of gastritis [p=.017] and gastric ulcer [p=.047]. Dyspepsia patients should be tested for cagA status along with the tests for H. pylori status; and a positive cagA testing should be considered as an indication for eradication of treatment

Role of urinary interleukin-8 in children with febrile urinary tract infection

lnterleukin-8 [IL-8] is produced within the urinary tract during urosepsis and is involved in the recruitment of neutrophils to the urinary compartment, in addition, deliberate colonization of the human urinary tract with E.coli resulted in a rapid increase in urine IL-8 levels without a detectable rise in IL-8 in serum. This study was conducted in Mansoura University Children's Hospital, and comprised 56 children with febrile urinary tract infection [UTI] and 10 matched healthy controls. Urine samples were taken from patients and control subjects and were analyzed by microscopic examination and bacterial culture. Urinary IL-8 measurements were performed for all samples. A significant increase in urinary IL-8 level was recorded in patients with febrile urinary tract infection compared to healthy controls [P<0.001]. The highest median urinary IL-8 level was found in febrile UTI with E.Coli, followed by staphylococcal infection. In conclusion, urinary IL-8 level may help in the diagnosis, and follow up of children with febrile UTIs