Detection of Neisseria gonorrhea, chlamydia trachomatis and mycoplasma genitalium in acute male urethritis using multiplex PCR

Acute urethritis is one of the most common STD Syndromes diagnosed in men. This study was to identify Neisseria Gonorrhea, Chlamydia trachomatis and Mycoplasma genitalium in acute male urethritis using Multiplex PCR technique and to compare urethral discharge versus urine as samples for diagnosis of urethritis. Evaluation of different microbiological techniques used for the diagnosis of gonococcal urethritis was also studied. Thirty adult males attending in the Venereal Disease Clinics, Ain Shams University Hospitals complaining of symptoms suggestive of acute urethritis were included in the study. Urethral discharge and first voided urine samples were subjected to wet and Gram stained smears, Gonococcal culture and Multiplex PCR for detection of Neisseria gonorrhea, Chlamydia trachomatis and Mycoplasma genitalium. Causative organisms of urethritis were identified in 22 out of the 30 studied patients [73.3%]. Gonococcal urethritis was diagnosed in 7 cases [23.3%], while non gonococcal urethritis due to Chlamydia trachomatis and Mycoplasma genitalium was detected in 10 [33.3%] cases. Simultaneous gonococcal and non gonococcal urethritis was identified in 5 cases [16.7%]. While the sensitivity of Gram stain for diagnosis of Gonococci was 75% while specificity was 100%. Culture, sensitivity and specificity were 58.2% and 100%, respectively, in comparison to the standard PCR test. Urethral discharge samples showed higher sensitivity compared to urine samples for detection of causative organisms of urethritis using PCR technique [78.9% Vs 45.4%, P

Porphyromonas gingivalis; prevalence and quantitation by pcr and clinical assessment of periodontal diseases in type I diabetes mellitus

The aim of this study is to assess the prevalence of periodontal diseases as well as dental caries in a group of diabetic children and adolescents and to study the subgingival microflora, Porphyromonas gingivalis [P.gingivalis]. The study was conducted on 70 type I diabetic patients, 20 healthy age-matched children and adolescents were included as a control group. Both patients and controls were subjected to periodontal examination and subgingival plaque samples for detection and quantitation of Porphyromonas gingivalis using PCR technique. Thirty eight among the seventy studied patients [54%] have been diagnosed as gingivitis [inflammation confined to the gingival], 9/70 [13%] as periodontitis [progressive destruction of periodontal ligament and alveolar bone with pocket formation], and the remaining 23 [33%] were periodontally healthy diabetics. A significantly higher percentage of Porphyromonas gingivalis positive PCR and higher DNA copies/ml were detected in periodontitis and gingivitis compared to periodontically healthy diabetics and healthy controls [P<0.05]. On comparing periodontitis and gingivitis groups, a statistically significant difference was detected [P<0.05] while periodontically healthy diabetics did not show any significant difference neither in positive PCR nor in DNA copies/ml compared to healthy controls [P<0.05]. Assessment of caries condition showed higher caries scores among diabetics than controls but this increase was statistically non significant [P>0.05]. A statistically significant difference was detected between age of the patient, disease duration, poor metabolic control and the development of periodontal disease. Periodontal diseases exist in a significant percentage of diabetic children and adolescents; higher age of the patient, longer duration of DM, and poor diabetic control are risk factors for the development of periodontitis and gingivitis; Porphyromonas gingivalis may be implicated in the development of periodontal disease. Detection and Quantitative analysis of this organism is important for the evaluation of periodontai diseases

Variations and genotypes of interleukin-10 in diabetes mellitus complicating chronic hepatitis C virus infection

To study whether genetic polymorphism influence lnterleukin-10 [IL-10] production and immune derangement that may contribute to the development of Diabetes Mellitus [DM] in chronic Hepatitis C Virus [HCV] infection, Two groups of HCV positive patients with liver cirrhosis [23 diabetic and 29 non-diabetic] were studied in addition to 10 healthy subjects. IL-10 serum levels were assayed for all the studied groups using ELISA technique. Two single nucleotide polymorphisms [SNPs] in the promoter region of IL-10 gene namely -1082 [A/G] and-592 [A/C] were genotyped in the HCV groups using Polymerase Chain Reaction, restriction fragment length polymorphism [PCR-RFLP] Technique. Serum IL-10 levels were found to be significantly higher in each of HCV groups compared to healthy control group [P<0.01] with positive correlation to liver enzymes, Fasting Blood Sugar [FBS] and negative correlation to serum albumin. Significant differences of IL-10 levels were detected in diabetic compared to non-diabetic HCV patients [9.94 +/- 3.5 versus 7.68 +/- 2.0 pg/ml, P<0.05].The frequency of carriage of allele G of-1082 A/G and allele C of-592 A/C markers were found to be higher in diabetic HCV compared with non diabetic patients [p=0.024, OR=3.12[95% Cl, 1.16- 8.39] and [p=0.045, OR=2.64 [95% Cl, 1.02, 6.84]] respectively. Haplotype analyses of both markers revealed that the carriage of haplotype GC was significantly higher in diabetic HCV compared to that of non diabetic patients [p<0.0001 and OR=7.49] [95% Cl, 3.45, 15.87]] It was concluded that IL-10 gene polymorphism and subsequently high IL-10 levels is associated with chronic HCV infection and may be involved the pathogenesis of diabetes mellitus

DNA Sequencing and bacieriophage based technique for rapid detection of rifampicin resistant mycobacterium tuberculosis

The drug resistant Mycobacterium tuberculosis [M TB] is an emerging problem of great importance to public health worldwide. This has stressed the need for development of more rapid techniques to accurately identify mycobacteria and drug resistant strains This study was done in an attempt to find a rapid and reliable method for identification of M. TB, and to study the genetic mutation in rifampicin resistance. Thirty sputum samples were subjected to identification of M. TB using conventional methods including Ziehli-Neelsen [ZN] stain and Lowenstein Jensen [LJ culture in comparison to the new rapid FAST plaque TB [bacteriophage based assay] which was used also to detect rifampicin resistance. The rifampicin resistant M. TB were subjected to PCR implication of rpoB gene followed by automated DNA sequencing. The phage technique identified 14/30 [46.7%] and when compared to LJ culture which was able to detect 19/30 [63.3%], the sensitivity was 73.7% and the specificity was 100%. Z.N. stain detected acid fast bacilli [AFB] in 10/30 ['33.3%]. Among the 20 smear negative samples the phage technique was able to detect 4 out of 9 cases identified by LJ culture with 44.4% sensitivity and 100% specificity. Among the rifampicin resistant M. TB, the highest percentage of point mutation was detected at codon 531[40%] and codon 526 [40%] followed by codon 516 [20%]. In conclusion the phage technique could be used in conjunction with sputum smear microscopy to detect additional cases which would be missed with smear alone. Rifampicin resistant M. TB is mostly associated with rpoB gene mutations

protective role of intestinal microflora [lactobacillus GG] in the pathogenesis of food allergic diseases

The aim of this study was to evaluate the role of intestinal microflora [Lactobacillus GG] in the development of food allergy in atopic patients. Sixty patients with atopic diseases due to food allergy [20 asthmatics, 20 with atopic skin diseases and 20 with allergic rhinitis] were examined for stool Lactobacillus GG and compared with 20 healthy controls. The results showed that atopic patients with food allergy in each group had a low stool Lactobacillus GG count as compared with the controls and there was an inverse correlation between stool Lactobacillus GG and both eosinophil count and serum IgE level in each group. It was concluded that there is a strong association between low Lactobacillus GG stool count and the occurrence of allergy in food allergic diseases

role of neuropeptidey in acute exacerbation of bronchial asthma in children

Neuropeptides are chemical mediators in the sensory parasympathetic and sympathetic neurons of the airways. Numerous neuropeptides are selectively and specifically released from the lung tissues e.g., neuropeptide Y [NPY], vasoactive intestinal peptide [VIP], calcitonin gene-related peptide [CGRP], substance P [SP] and tachykinins [TK]. The imbalance between these neuropeptides is considered as one of the underlying mechanisms of bronchial asthma as some of them are potent bronchodilators whereas others are potent bronchoconstrictors. NPY is one of the important neuropeptides involved in the regulation of mechanisms of bronchial asthma. The aim of this work is to study the role of NPY in acute exacerbation of bronchial asthma in children. The study included two groups: patient group consisting of 30 children selected on clinical basis being presented with acute exacerbation of bronchial asthma and 20 apparently healthy age and sex-matched children as controls. None of the patients used oral steroids. For all patients, two plasma samples were obtained before and after treatment with bronchodilators whereas one plasma sample was collected from each member of the control group. Plasma level of NPY was determined by quantitative enzyme immunoassay after an extraction step for the tested plasma by column extraction method to separate peptides from any interfering substance. There was a significant higher plasma levels of NPY for patients in comparison to controls [10.5 +/- 4.6ng/ml versus 0.13 +/- 0.05 ng/ml P < 0.001] whereas there was no significant difference between NPY plasma levels before and after treatment [10.5 +/- 4.6ng/ml versus 10.02 +/- 4.1ng/ml; P>0.05]. The increased plasma levels of NPY during exacerbation of bronchial asthma act as a breakdown mechanism against the exacerbation of this asthma so it can be useful as a diagnostic tool for asthma but can't be employed as a predictor for responsiveness to therapy like other neuropeptides as there was no significant difference between its levels before and after treatment. Further future studies are recommended for better underst and ing of the neural mechanisms of NPY and other neuropeptides in bronchial asthma which might provide. fruitful therapeutic approach