Detection of west nile virus nucleic acid in egyptian blood donors by real time PCR

Transfusion-transmitted West Nile virus [WNV] was first reported in 2002 led to rapid development of investigational nucleic acid tests [NAT]. This study aimed to screen WNV by real time RT-PCR among apparently healthy Egyptian blood donors. A total of 284 volunteer blood donors [197 males and 87 females] were enrolled in this study. Blood samples [5ml] were collected in sterile vacutainer tubes containing EDTA under aseptic conditions. After centrifugation plasma were separated and collected into two tubes, one used for routine screening tests for donated blood. From the second tube, 200micro L plasma from 6 samples were pooled into minipools automatically and used for screening of WNV by real-time RT-PCR while the remaining two samples were tested individually. In positive reactive minipools to WNV, retesting individual samples were performed. Out of the 47 pools and 2 individual samples tested for WNV, 7 minipools were positive. These 7 minipools contain 42 individual samples and after retesting them, 7 [2.5%] samples were positive for WNV. Routine screening of blood donors revealed 7 positive samples for HBsAg. 11 positive samples for HCV Abs [3.9%], two samples [0.7] were positive for HCV RNA by PCR and 5 [1.7%] positive samples of syphilis tested by Treponema pallidum haemagglutination assay [TPHA]. WNV can be transmitted through blood transfusion in Egypt and to overcome this problem screening of blood donors against WNV should be implemented in Egypt according to CDC recommendation

CX[3]C Chemokine fractalkine and IL-18 in allergic bronchial asthma

Human bronchial asthma is characterized by airway hyper responsiveness [AHR], eosinophilic airway inflammation, mucous hypersecretion and high serum level of IgE. AHR and mucus over secretion are often linked to asthma symptoms and morbidity. Although the mechanisms underlying these features are complex, it is widely accepted that TH2 cells which produce a limited repertoire of cytokines are responsible for inducing these characteristic features of bronchial asthma. The aim of this study is to investigate the levels of serum soluble fractalkine [CX3C] and interleukin IL-18 in a group of patients with bronchial asthma. This study was conducted on 55 subjects who were divided into three groups, 20 patients with bronchial asthma who were not receiving corticosteroidal treatment, 20 patients with bronchial asthma and were receiving inhaled steroids such as beclomethasone diproprionate and 15 normal subjects as a control group. The patients were selected from either inpatients in Al-Zahraa University Hospital internal medicine or outpatients from the internal medicine Clinic. Five milliliters of peripheral venous blood were collected and divided into two tubes; one of them contained EDTA to be tested for complete and differential blood count. From the other tube, serum is separated and stored at - 80?C until serum levels of soluble fractalkine, IL-18 and IgE are estimated using commercially available ELISA kits. In the present study serum soluble fractalkine [3535.3 +/- 3697.3] pg/ml, IL-18 [1431.5 +/- 716.58] pg/ml and serum IgE[257.27 +/- 73.300] IU/ml levels were higher in asthmatic patients than healthy controls which were [72.000 +/- 7.270] pg/ml, [352.86 +/- 127.29] pg/ml and [63.573 +/- 19.719] IU/ml respectively with statistical significant difference between them. By comparing these parameters between group1 and group 2, it was found that serum soluble fractalkine level was higher in group 2 [6700.0 +/- 2054.8] pg/ml than group 1 [170.50 +/- 57.260] pg/ml and the difference between them was highly significant. On the other hand IL-18 [1362.8 +/- 654.03] pg/ml was slightly lower in group 2 than group 1[1500.3 +/- 785.05] pg/ml and the statistical difference between them were not significant. Serum IgE level was lower in group 2 [231.75 +/- 80.733] IU/ml than group 1[282.79 +/- 56.030] IU/ml with statistically significant difference between them. In conclusion, both serum IL-18 and soluble fractalkine were elevated with allergic bronchial asthma independently of serum IgE level. Furthermore, corticosteroid inhalation has no effect on IL-18. On the other hand, it decreases the level of serum IgE but also increases the level of serum soluble fractalkine

Detection of Panton-valentine Leukocidin gene by real time polymerase chain reaction in community-acquired methicillin resistant staphylococcus aureus

Panton-Valentine leukocidin [PVL] is a biocomponent cytotoxin and assumed S. aureus virulence factor that has been associated with skin and soft tissue infections together with more severe manifestations including necrotising pneumonia. PVL has been strongly associated with emergence of community acquired methicillin-resistant S. aureus [CA-MRSA]. Objective: The aim of the current study is to determine whether carriage of PVL could be used as surrogate marker for CA-MRSA strains using rapidreal time PCR. Methods: The study was conducted on 92 community-acquired Staphylococcus aureus isolates which caused different types of infections and isolated from inpatients and outpatients attending different departments at Al-Zahraa Hospital. All isolates were identified by colony morphology on mannitol salt agar medium, Gram stain, positive reaction for catalase, coagulase and DNAase. Biochemical and antibiogram susceptibility testing were performed using walkaway MIC plates. MRSA strains were tested for penicillin binding protein 2a [PBP2a] using Mastalex Kit and PVL gene by real time polymerase chain reaction [RT-PCR] . Results: Out of 92 community-acquired Staphylococcus aureus isolates 34 [36.96%] were MSSA, while 58 [63.04%] were MRSA strains. Detection of PBP2a by Mastalex Kit was false negative in 6 [10.3%] of MRSA isolates. Detection of PVL gene by real time PCR was positive in only 23 isolates [39.7%]. Antimicrobial susceptibility testing revealed that all MRSA strains were resistant to lactam, cephalosporin, Carbapenems antibiotics and 91.4% of MRSA strains were resistant to tetracycline, while 70.7% were resistant to aminoglycosides and 5.2% were resistant to vancomycin. In Conclusion: During recent years, methicillin-resistant Staphylococcus aureus continues to be a major cause of both health care-associated and community-associated infections. Rapid and reliable detection with implementation of infection control measures is essential in limiting the spread of this organism. In this study, it was found that carriage of PVL cannot be used as a sole marker for CAMRSA and further studies are required to find a reliable marker or combination of markers to facilitate the recognition of CA-MRSA strains

study of some immunological indicators in children with down's syndrome

Abstract: Down's syndrome [DS] is the most common chromosomal abnormality in humans. It is characterized by precocious immunologic aging that results, among other things, in alterations of T lymphocyte subsets and natural killer cells. DS subjects have an increased prevalence of autoimmune disorders. The most common autoimmune disease in DS is related to the thyroid gland

Aim of work: investigate some indicators of immune response, such as white blood cells, lymphocytes, CD3+, CD4+, CD8+, CD56+ peripheral blood mononuclear cells and to study thyroid function and the presence of thyroid autoantibodies in DS children and to compare them with the same items in healthy children

Subjects and Methods: This study was carried on twenty six children; 15 children with DS, their age ranged from 13 to 63 months [32.8+/-15.6] and 11 healthy children matching in age and sex with DS children. Five ml venous blood samples were collected aseptically from each child. Evaluation of total leucocytic count, lymphocytes, CD3+, CD4+, CD8+ and CD56+ cells was carried for each subject. TSH and FT4 were measured by chemiluminesence and thyroid autoantibodies [antithyroglobulin and anti TSH receptor antibodies] were measured by radioimmunoassay

Results: Total leucocytic count, Lymphocyte count, CD3+ and CD4+ cells were lower in DS children [total leucocytic count was 5773.3 cells /mm3 +/- 1862.2, lymphocyte count was 2235.3+/-598.9, CD3+ cells were 1775.3+/-397.6, and CD4+ cells were 762.0+/-299.5] than healthy controls [total leucocytic count was 7909.1+/-1465.9, lymphocyte count was 3160.0+/-723.6, CD3+ cells were 2253.1+/-637.9, CD4+ cells were 1390.4+/-380.5] with statistical significant difference. CD8+ and CD56+ cells were higher in DS children [CD8+ cells were 980.5+/-286.3 and CD56+ cells were 394.3+/- 104.0] than healthy controls [CD8+ cells were 742.9+/-171.7 and CD56+ cells were 176.6+/-53.9] with statistical significant difference. CD4/CD8 ratio was reversed in DS children [0.79+/- 0.28]. Seven patients [46.7%] with DS were hypothyroid according to thyroid profile tests; while none of the control group revealed any abnormality in thyroid function. Thyroid autoantibodies were detected in 3 [20%] of DS children [2 hypothyroid and 1 euthyroid patients] and in none of the control group. In conclusion, a complex impairment of T-lymphocytes and natural killer cell production takes place in DS children. The sum of these alterations is likely the cellular basis of the defective immune responses and of the increased susceptibility to infectious diseases occurring in DS subjects. Thyroid dysfunction is common in DS children and thyroid autoantibodies are not uncommon in preschool DS children