Association of immune responses to mycobacterium tuberculosis peptide antigens with host genetic factors

Apolymerase chain reaction [PCR], based on insertion sequence IS6110,was developed to detect mycobacterium tuberculosis complex organisms in the blood samples of 56 tuberculosis patients and 34 healthy controls.the erly secreted antigenic target 6-KDa[ESAT-6] are used to stimulate T lymphocyte subsets from tuberculosis infected patients and the correltion of these immune responses to the genetic factoes[HLATYPE] which determined the host immune response is evaluated.ESAT-6 derived peptides:P1[1.05 +/- 0.084],P2[1.08 +/- 0.094],P3[1.02 +/- 0.086],P5[0.98 +/- 0.117] and P7[1.26 +/- 0.152] were significantly higer in the infected group than in non-infected one. Besides, 33ptients and 12 controls were tested for HL-DRB HLA DQB and HLA-DPB Only type HLA-DEB 15was significan tly associated with tuberculosis infection using the Chi-square test[X[2]=0.04311]. By using the relative risk some HL types were relatively more susceptible to be associated with tuberculosis infection. HLA-DR typing of patients showed that they covered a large spectrum of HLA-DR molecules encoded by HLA-DEB1,-DRB3,-DRB4,and DRB5genes.however,HLA-DQ typing showed that they HLA-DQB1 molecules, HLA-DP typing of patients showed that covered a large spectrum of HLA-DP molecules encoded by HLA-DPB1.

Candidemia in neonates

Neonatal fungal sepsis is a risky condition that needs effort for early diagnosis and management Clinical diagnosis of sepsis is not easy, because symptoms and signs are not specific, and complication of clinical conditions can occur rapidly, even in asymptomatic newborn infants, long before blood culture results are available. The aim of this work was to study risk factors for neonatal candidemia and evaluate the role of molecular biology in rapid diagnosis of neonatal sepsis using Polymerase Chain Reaction technique. One hundred neonates were studied. Twelve cases showed infection with yeast species [12% of neonates]. Three yeast species were isolated from blood of neonates. Candida albicans was the predominant species, and was isolated from 10 cases representing 83.3% of the total yeast isolates, while Non albicans Candida was isolated from two cases only, representing 16.7% of the total yeast isolates. They were identified as Candida parapsilosis and Debaryomyces hansenii which also known as Candida famata. Low birth weight and preterm delivery were significantly increasing the incidence of neonatal fungal sepsis. Detection of candidemia in neonatal blood was carried out by polymerase chain reaction [PCR] for universal yeast to amplify the V3 region of the large subunit ribosomal DNA. Analysis of neonatal cases showed that there is a perfect correlation between molecular and microbiological data in all infected cases. This means that PCR for universal yeast is recommended to be used for confirmation of fungemia in neonates, long before blood culture results are available

Detection of microdeletions involving the DAZ locus in idiopathic male infertility

Deletions of the AZFc [azoospermic factor c] region of the Y chromosome including DAZ gene are the most common known cause of spermatogenic failure. This study was conducted with the aim of detecting Y chromosome microdeletions involving the DAZ locus in idiopathic male infertility to allow rapid and accurate diagnosis required for proper genetic counseling. The study included 30 male patients with idiopathic azoospermia [24/30] or oligozoospermia [6/30]. A control group consisted of 10 normal fertile males and 5 females. All cases were subjected to detailed history, clinical examination, assessment of testicular volume, semen analysis, serum hormonal profile [FSH, LH, testosterone], testicular biopsy, chromosome analysis, polymerase chain reaction [PCR] amplification of two specific loci of the DAZ gene on the Y chromosome [sY254 and sY255], single strand conformations polymorphism analysis [SSCP]. The result of the study revealed that deletions involving the sY245 and sY255 DAZ loci were found in 4 cases [4/30; 13.3%]; 3 azoospermic patients and one with severe oligozoospermia. All four cases with microdeletions had decreased testicular volume, normal serum LH and T, serum FSH was elevated in 3 of them and normal in one. The two loci were amplified normally in the male control group and failed to amplify in the female control group. SSCP analysis failed to find any point mutations in sY254 and sY255 in patients with absence of microdeletions of DAZ gene. In conclusion, the estimated frequency of microdeletions involving the DAZ locus is 13.3% in azoospermic and severely oligozoospermic Egyptian men with idiopathic infertility. Polymerase chain reaction amplification of the DAZ locus is a rapid and accurate method for the diagnosis of microdeletions of the Y chromosome in patients with idiopathic infertility especially those seeking micromanipulation assisted reproduction

Polymerase chain reaction in pulmonary tuberculosis: Sputum versus blood

- The aim of this work is to study the role of polymerase chain reaction [PCR] in the diagnosis of pulmonary tuberculosis and to compare the sensitiviity of its utility in sputum and blood of patients with pulmonary tuberculosis. The present study entailed 25 patients with recently diagnosed pulmonary tuberculosis. All patients were subjected to sputum culture on Lownstein Jensen Medium. Radiological investigation, PCR assay for deteection of mycobacterial DNA in sputum and peripheral blood. The current study revealed that sputum PCR assay for mycobacterium tuberculosis was positive in 24 patients out of 25 patients positive ZN stain and culture with sensitivity 96%. As regard blood PCR assay comparing to positive sputum culture. 20 cases were positive out of 25 positive cases by ZN stain and culture with sensitive 80%. The sensitivity of blood PCR as compared sputum PCR was 80.33% while specificity of blood PCR to sputum PCR was 100%. Sputum PCR was found to be a rapid and sensitive technique for the diagnosis of pulmonary tuberculosis. Blood PCR in patients with mycobacterium tuberculosis may be considered as an additional effective diagnosis tool with results comparable with sputum PCR

Detection of hepatitis E virus in greater Cairo two wastewater treatment plants and its prevalence among workers of these plants

Because of inadequate public sanitation, epidemics of HEV infection have been reported in several developing countries. HEV-specific cDNA was prepared by reverse transcription of the total RNA extracted from water samples. Specified DNA amplification by PCR demonstrates the presence of HEV in sewage samples from the inlets [PCR positive 10/36:27.77%], while outlet samples were PCR negative. HEV IgM was detected in 40 workers out of 78 in these two studied plants [age 20-60 years], with a percentage of 51.25% and HAV IgM was also detected in 3 workers out of 78 [3.84%]. The study of serum ALT and albumin level go with the increase in HEV IgM in sera. This study which was carried out in two different wastewater treatment plants showed that HEV contamination was higher in one of them [El-Berka] than El-Zenin. A total of 33.33% of influent water samples were positive and 55.25% of workers were HEV IgM positive in the first plant [E1-Berka] and the corresponding figure in the other plant [El-Zenin] showed lower contamination, 22.22% in influent water sample and 40.9% of workers