Prevalence of hepatitis C virus among HIV-infected patients.

BACKGROUND AND OBJECTIVES: Hepatitis C virus and Human Immunodeficiency Virus (HIV) share the same rate of transmission. HIV/HCV co-infected individuals may result in faster progression of liver fibrosis and highly increase the risk of cirrhosis, hepatocellular carcinoma development. Thus this study was conducted to determine co-infection of HCV genotypes in positive HIV patients in Ahvaz city, Iran. MATERIALS AND METHODS: The sera samples were collected from confirmed 78 infected HIV, 67 (85.89%) males and 11 (14.1%) females. All sera samples were tested for HCV Ab using ELISA test. The HCV Ab positive samples were tested for detection of 5' untranslated (UTR) and core regions of HCV genome using nested RT-PCR. The PCR products of 5UTR and core regions were sequenced to determine HCV genotypes. RESULTS: Among the 78 infected HIV, 25 (32.05%) cases including 20 (25.64%) males and 5 (6.41%) females were positive for HCV Ab (p=0.316). 53 (67.94%) of HIV patients were negative for HCV Ab. Among 25 positive HCV Ab, 19 (24.35%) cases including 15 (19.23%) males and 4 (5.12%) females were positive for HCV RNA (p=0.447). The PCR products of 5 positive samples were randomly sequenced. The results of sequences and alignments showed that the detected HCV genotypes were three 3a and two 1a. The occurrence of genotype HCV 1a was found in one male injecting drug user Injecting Drug User (IDU) and one female. The HCV 3a genotype was detected in the three males IDU. CONCLUSION: The results of this survey indicated that 32.05% of HIV patients were positive for HCV Ab, among them 24.35% were positive HCV RNA. HCV genotype 3a was dominant and detected in the three males IDU. Regarding the consequences of HIV/HCV co-infection, it is suggested that HCV RNA detection should be regularly checked in individuals infected with HIV.

Prevalence of Hepatitis G Virus Among Hemodialysis and Kidney Transplant Patients in Khuzestan Province, Iran.

BACKGROUND: Hepatitis G virus (HGV) is a member of Flaviviridae. Prevalence of HGV in healthy people is very low, but this virus is more prevalent in patients with hepatitis. Besides, relative frequency of HGV in patients undergoing hemodialysis, and kidney recipients is very high. The role of HGV in pathogenesis is not clear. Since this virus cannot be cultivated, molecular techniques such as Revers Transcription Polymerase Chain Reaction (RT-PCR) is applied to detect HGV. OBJECTIVES: The current study aimed to investigate the prevalence of HGV using determination of E2, viral envelope antigen, antibodies and the RNA by Enzyme Linked Immunosorbent Assay (ELISA) and RT-PCR techniques. The rational of the study was to determine the prevalence of HGV in patients undergoing hemodialysis and kidney transplantation in Khuzestan province, Iran. PATIENTS AND METHODS: Five hundred and sixteen serum samples of the patients undergoing hemodialysis and kidney transplantation from various cities of Khuzestan province were collected. Anti-hepatitis G E2 antibodies were investigated by ELISA method. RNAs were extracted from serums and Hepatitis G RNA was detected by RT-PCR. RESULTS: Of the 516 samples, 38 (7.36%) specimens were positive for anti-HGV by ELISA. All of these ELISA positive samples were negative for HGV genome by RT-PCR. Of the remaining 478 ELISA negative samples, 16 (3.14%) samples were positive by RT-PCR. CONCLUSIONS: Hepatitis G Virus was not prevalent in the patients undergoing hemodialysis and kidney transplantation in Khuzestan province. Although reports indicated high frequency of co-infection of HGV with hepatitis B and C viruses, in the current research, co-infection of HGV with B and C was not considerable. Since different groups and subtypes of HGV are reported, periodic epidemiologic evaluation of HGV and its co-infection with other hepatitis viruses is suggested in other populations such as the patients with thalassemia; however, periodic epidemiologic monitoring of HGV may be helpful to control future potential variations of the virus.

Hepatitis B virus Genotyping Among Patients With Cirrhosis.

BACKGROUND: Hepatitis B virus (HBV) infection is a worldwide public health problem. Nine HBV genotypes (A-I) have been already discovered. HBV genotypes are important both in the clinical manifestation of disease and treatment response. Moreover, HBV DNA without HBs (Hepatitis B surface)-antigenemia was detected in some patients with chronic hepatitis (occult hepatitis). There is little information about HBV genotypes and its relation to occult infection despite the importance of this infection in Khuzestan Province. OBJECTIVES: This study aimed to determine both occult hepatitis B infection and HBV genotypes among cirrhotic patients. PATIENTS AND METHODS: Thirty-eight patients with liver cirrhosis, including 11 (28.9%) HBsAg-positive patients and 27 (71.1%) patients with cryptogenic cirrhosis participated in this study. The mean age of the patients at the time of cirrhosis diagnosis was 54.85 years (range 26-75 years). All patients were anti-HCV and anti-HIV negative. For all the samples, the serological Enzyme-Linked Immunosorbent Assay (ELISA) was performed for HBV markers including HBsAg, HBcAb, HBeAg, HBeAb tests. The common primer of S region of HBV was used for Nested PCR. The PCR products of the positive individuals were sequenced for genotyping and subtyping of HBV. RESULTS: Eleven (40.7%) out of 27 HBV cryptogenic cirrhosis and all 11 HBsAg-positive patients were positive for HBV DNA. The seroprevalences of Hepatitis B virus HBe antigen, anti-HBe and anti-HBc antibodies among the cryptogenic cirrhosis patients were 5 (18.5%), 1 (3.7%), and 5 (20.83), and among HBsAg-positive patients were 6 (54.5%), 5 (45.5%), and 7 (63.6%), respectively. CONCLUSIONS: In our study, only HBV genotype D was found among all the positive HBsAg and occult HBV infection. Moreover, high prevalence (40.7%) of occult HBV infection was determined among patients suffered from cryptogenic cirrhosis.

Bioconjugated fluorescent silica nanoparticles for the rapid detection of Entamoeba histolytica.

Rapid detection of Entamoeba histolytica based on fluorescent silica nanoparticle (FSNP) indirect immunofluorescence microscopy was evaluated. Silica nanoparticles were synthesized using Stöber's method, with their surface activated to covalently bind to, and immobilize, protein A. For biolabeling, FSNP was added to conjugated E. histolytica trophozoites with monoclonal anti-E. histolytica IgG1 for microscopic observation of fluorescence. Fluorescent silica nanoparticle sensitivity was determined with axenically cultured E. histolytica serially diluted to seven concentrations. Specificity was evaluated using other intestinal protozoa. Fluorescent silica nanoparticles detected E. histolytica at the lowest tested concentration with no cross-reaction with Entamoeba dispar, Entamoeba moshkovskii, Blastocystis sp., or Giardia lamblia. Visualization of E. histolytica trophozoites with anti-E. histolytica antibody labeled with fluorescein isothiocyanate (FITC) was compared with that using anti-E. histolytica antibody bioconjugated FSNP. Although FITC and FSNP produced similar results, the amount of specific antibody required for FITC to induce fluorescence of similar intensity was fivefold that for FSNP. Fluorescent silica nanoparticles delivered a rapid, simple, cost-effective, and highly sensitive and specific method of detecting E. histolytica. Further study is needed before introducing FSNP for laboratory diagnosis of amoebiasis.

The Prevalence Rate of Helicobacter pylori Infection in, Chronic Otitis Media With Effusion Patients.

BACKGROUND: Otitis media with effusion (OME) is a common disease in children. Viral or bacterial infections, allergy, adenoids, functional abnormalities of the Eustachian tube, and gastroesophageal reflux might have a possible role in the pathogenesis of OME. However, the exact pathogenesis of OME is still unsettled. OBJECTIVES: The purpose of this study was to compare Helicobacter pylori prevalence rates in the nasopharynx of pediatric patients with and without OME. PATIENTS AND METHODS: Eighty-four patients (50 males and 34 females) who were subjected to adenoidectomy and myringotomy were included in the study group. Ninety-one patients (48 males and 43 females) who had only adenoidectomy were selected as the control group. Detection of H. pylori was done by polymerase chain reaction (PCR). RESULTS: Adenoid samples were positive for H. pylori in 21 (25%) patients in study group and 18 (19.8%) patients in control group. In the study group, 36 (42.8%) effusion samples (otitis media) of the patients were positive for H. pylori. In an analysis that compared H. pylori-negative and -positive children, the odds ratio (OR) for the occurrence of H. pylori was 1.35 (95% CI, 0.66 - 2.71). The association of age with H. pylori positivity decreased for 1-5 years age group, (1.09; 95% CI, 0.39 - 3.05) but increased for the 6-10 years group (OR, 1.48; 95% CI, 0.61-3.58). Furthermore, the association of sex with H. pylori positivity decreased for the male group (OR, 1.21; 95% CI, 0.50 - 2.91), but increased in the female group (OR, 1.44; 95% CI, 0.51-0.4.05). CONCLUSIONS: Heavy colonization of H. pylori in adenoid tissue and middle ear might have a role in pathogenesis of this infection. For OME cases resistant to medical treatment, it might be meaningful to evaluate the patient for H. pylori.

Protective Effects of Crocin on Ischemia-reperfusion Induced Oxidative Stress in Comparison With Vitamin E in Isolated Rat Hearts.

BACKGROUND: Myocardial Injury caused by ischemia-reperfusion leads to cardiac dysfunction, tissue injury and metabolic changes. The production of reactive oxygen species (ROS) and lipid peroxidation are accompanied by ischemia-reperfusion injury. OBJECTIVES: The aim of this study was to assess the cardio protective potential effects of crocin in comparison with vitamin E on antioxidant capacity in ischemia-reperfusion of isolated rat hearts. MATERIALS AND METHODS: Seventy male Sprague-Dawley rats were randomly divided into seven groups, including: sham, control and experimental groups treated with different doses of crocin (10, 20 and 40 mg/kg) or vitamin E (100 mg/kg) and a combination of crocin (40 mg/kg) with vitamin E (100 mg/kg) that were administrated orally for 21 days. The heart was quickly excised, transferred to a Langendorff apparatus at constant pressure and subjected to 30 minutes of global ischemia followed by 60 minutes of reperfusion. Cardiac damage markers and antioxidant enzymes were measured. RESULTS: The results showed that superoxide dismutase and catalase enzyme activities increased and Mallon de aldehyde (MDA) decreased in animals pretreated by crocin (40 mg/kg) and vitamin E (100 mg/kg). Moreover, there was a significant improvement in post ischemic recovery of antioxidant capacity during reperfusion in rats receiving a combination of crocin (40 mg/kg) and vitamin E (100 mg/kg). CONCLUSIONS: The results demonstrated the protective role of crocin on antioxidant capacity, which may partially be related to stability or amplification of antioxidant systems. Like vitamin E, crocin may be beneficial for prevention or treatment of cardiac dysfunction and myocardial infarction in patients with ischemic heart disease.

Nocardia co-infection in patients with pulmonary tuberculosis.

BACKGROUND: Tuberculosis (TB) remains as one of the most serious infectious diseases in the world. Pulmonary tuberculosis can occur with other pulmonary diseases caused by opportunistic organisms such as Nocardia spp. particularly in immunocompromised patients. Therefore, diagnosis of co-infection at the early stage of the disease could be lifesaving. OBJECTIVES: The goal of this study was to detect Mycobacterium tuberculosis and Nocardia spp. in sputum specimens in order to assess the concomitant nocardiosis and tuberculosis in patients with suspected pulmonary tuberculosis. PATIENTS AND METHODS: From March 2011 to April 2012, 189 sputum specimens were obtained from patients who were suspected of having pulmonary tuberculosis. Out of 189 samples, 32 of the samples belonged to hospitalized HIV-infected patients. Samples were examined by Gram and Ziehl-Nelsen staining, culture and PCR methods. RESULTS: From 157 sputum specimens, positive samples by acid fast staining, culture and PCR for M. tuberculosis were reported for 7.6% (12/157), 10.1% (16/157) and 7% (11/157) of samples, respectively. No results were obtained by the described methods for Nocardia spp. Among 32 samples of HIV-infected patients, four (12.5%) had positive results for acid fast staining, culture and PCR detecting M. tuberculosis while only two samples had positive results for Nocardia spp. by PCR and no results were reported by culture, Gram and acid fast staining for this organism. CONCLUSIONS: Concurrent pulmonary nocardiosis and tuberculosis is frequent in HIV-infected patients. Rapid and sensitive methods such as PCR are recommended for detection of such co-infections.

Validity of bioconjugated silica nanoparticles in comparison with direct smear, culture, and polymerase chain reaction for detection of Mycobacterium tuberculosis in sputum specimens.

BACKGROUND: Tuberculosis is a public health problem worldwide, and new easy to perform diagnostic methods with high accuracy are necessary for optimal control of the disease. Recently, fluorescent silica nanoparticles (FSNP) has attracted immense interest for the detection of pathogenic microorganisms. The aim of this study was to detect Mycobacterium tuberculosis in clinical samples using bioconjugated FSNP compared with microscopic examination, polymerase chain reaction (PCR), nested PCR, and culture as the gold standard. METHODS: In total, 152 sputum specimens were obtained from patients who were suspected to have pulmonary tuberculosis. All samples were examined by the four techniques described. RESULTS: The assay showed 97.1% sensitivity (95% confidence interval [CI] 91-99.2) and 91.35% specificity (CI 78.3-97.1). Furthermore, assays using variable bacterial concentrations indicated that 100 colony forming units/mL of M. tuberculosis could be detected. There were no differences between the results obtained from two types of mouse monoclonal antibody against Hsp-65 and 16 KDa antigens. CONCLUSION: We performed this assay in a large number of clinical samples to confirm the diagnostic specificity and sensitivity of the test and can recommend its application for diagnosis of M. tuberculosis. We believe that this method is more convenient for routine diagnosis of M. tuberculosis in sputum and will be more easily applicable in the field, and with sufficient sensitivity.

Comparison of fluorescent in situ hybridization and histological method for the diagnosis of Helicobacter pylori in gastric biopsy samples.

BACKGROUND: The establishment of Helicobacter pylori in the stomach and duodenum is associated with gastritis, peptic ulcers, and gastric cancer. Application of suitable methods, including molecular techniques, for an accurate detection of H. pylori can lead to the administration of appropriate drugs and successful therapy. In this study, fluorescent in situ hybridization (FISH) was compared with histology for the diagnosis of H. pylori in gastric biopsy specimens. MATERIAL/METHODS: Flourescently labeled oligonucleotdie probes that target ribosomal RNA were utilized in the FISH procedure. Ninety-one gastric biopsy specimens were tested by FISH and by histology using hematoxylin-eosin (H-E) and Geimsa stains. Furthermore, clarithromycin resistance in 39 of the 91 specimens was examined by FISH. RESULTS: The sensitivity and specificity of FISH for the detection of H. pylori were 97.9% and 100%, respectively. Of the 39 samples that were tested for clarithromycin resistance, 19 were FISH positive for H. pylori, of which 15 and 4 specimens were infected with clarithromycin-susceptible and clarithromycin-resistant strains, respectively. There were coccoid forms of H. pylori in a few of the specimens. CONCLUSIONS: FISH is a highly sensitive and specific technique for the diagnosis of H. pylori infection. It can be a method of identification when a patient is infected with coccoid forms of H. pylori. The ability of FISH for determination of clarithromycin resistance is a considerable advantage of this method over histology.

Rapid detection of clarithromycin-resistant Helicobacter pylori in patients with dyspepsia by fluorescent in situ hybridization (FISH) compared with the E-test.

BACKGROUND: Clarithromycin is the antibiotic of choice for treatment of H.pylori-related dyspepsia, but unfortunately, resistance to clarithromycin is not rare. Detection of resistant strains takes 2 to 4 days by conventional methods. In this report, we applied the FISH technique for rapid detection of H.pylori in biopsies of dyspeptic patients. METHODS: Gastric biopsies from 50 patients suffering from dyspepsia were tested in this study. Part of each biopsy specimen was cultured and the remainder was fixed in liquid nitrogen. After mounting of frozen sections on microscopic slides, they were hybridized with oligonucleotide probes for detection of clarithromycin-resistant H.pylori. The slides were visualized under a fluorescent microscope. Susceptibility of cultured strains of H. pylori to clarithromycin was also determined by the E-test and the results were compared. RESULTS: Twenty-five of 50 biopsy specimens examined by FISH were positive for H.pylori. FISH showed that 17 strains (68%) were susceptible to clarithromycin and 6 strains (24%) were resistant. Bacteria isolated following culture of 2 biopsy specimens had a mixture of both clarithromycin-susceptible and resistant strains (8%). There was no discrepancy between the E-test and FISH technique for detection of resistant strains of H.pylori. CONCLUSION: FISH is a rapid technique for detection of H.pylori in clinical samples. Moreover, strains susceptible to clarithromycin can be detected quickly. Therefore, this method is suitable for determination of susceptibility of H.pylori to clarithromycin, especially when a quick decision is necessary for treating dyspeptic patients.

Application of fluorescent in situ hybridization (FISH) for the detection of Helicobacter pylori.

BACKGROUND: Peptic ulceration following infection of the stomach with H. Pylori is a common disease. Accurate and rapid detection of the bacteria can lead to the implementation of appropriate treatment and recovery. Chronic infection of the gastric milieu with H. Pylori may lead to gastric carcinoma. Routine detection of this bacterium in peptic ulcer is based on the urease test and culture of peptic biopsies. Unfortunately, the sensitivity and specificity of both tests are not satisfying. Molecular techniques have been successfully applied for the rapid and accurate detection of bacterial agents in clinical samples. This study was undertaken to evaluate the sensitivity and specificity of fluorescent in situ hybridization (FISH) in the detection of H. Pylori in patients suffering from dyspepsia. MATERIAL/METHODS: One hundred gastric biopsy samples taken by endoscopy from the antrum and corpus of the stomach were tested by FISH and compared with the conventional culture method complemented by biochemical tests. RESULTS: FISH detected H. Pylori in 48 clinical samples, while the conventional method detected 42 samples. The sensitivity and specificity of FISH for the detection of H. Pylori were calculated as 98% and 100%, respectively. CONCLUSIONS: The findings of this study suggest that FISH is a highly suitable and rapid method for diagnosing H. Pylori. Especially when the samples are taken from the antrum and the corpus of the stomach, this technique potentially can be applied routinely for the detection of this bacterium in clinical samples.