Open Access Original Research Article

Development of a New Bifunctional Fusion Protein of Vaccine Strains Clostridium perfringens Type D and Clostridium septicum Epsilon-alpha Toxin Genes

Marzieh Kamalirousta, Reza Pilehchian

Microbiology Research Journal International, Page 1-9
DOI: 10.9734/MRJI/2018/39261

There are various difficulties regarding of produce and design of a suitable fusion protein, and functionality is the most important of this problem. For producing a functional fusion protein, it is necessary to have a proper suitable host to clone a properly designed fusion gene. Clostridium perfringens and C. septicum are important pathogens of humans and livestock and produce numerous toxins including epsilon and alpha, which are responsible for severe diseases. In this study, a new construct of C. perfringens type D epsilon and C. septicum vaccine strains alpha toxin genes designed in addition, cloned into a prokaryotic host. Online software used for in silico, study and the prediction of the fusion protein construct primary, secondary and tertiary structures. At the next step, specific primers used for amplification of these genes, based on nucleotide sequences that retrieved from GenBank. The amplified etx and CSA fragments linked together by a linker based on a helix forming peptide (i+4) E, K. The linker introduced between two domains by fusion PCR and using Pfu DNA polymerase, epsilon forward and alpha reverse primers. The fusion gene ligated into pGEM-B1cloning vector and cloned into E. coli strain TOP10. This epsilon-alpha fusion gene could be used for development of a recombinant epsilon alpha fusion protein vaccine.

 

Open Access Original Research Article

Evaluation of Xpert MTB/ RIF Assay for the Detection of Female Genital Tuberculosis in a Tertiary Care Center- A Descriptive Cross-sectional Study

Anjum Farhana, Danish Zahoor, Munazah Manzoor, Farhath Kanth

Microbiology Research Journal International, Page 1-6
DOI: 10.9734/MRJI/2018/39636

Aims: Female genital tuberculosis (FGTB) is a silent disease and patients may present with non-specific symptoms irrespective of disease severity and duration. It is a major cause of severe tubal disease leading to infertility. A high degree of suspicion is required for its diagnosis. Conventional diagnostic tests have low sensitivity due to the paucibacillary nature of FGTB. Polymerase chain reaction (PCR) based methods like Xpert MTB/RIF assay hold potential for the early diagnosis of FGTB. The objective of this study was to evaluate the utility of Xpert MTB/RIF assay for the diagnosis of FGTB.

Methodology: Laparoscopy directed endometrial biopsies of 87 samples of females suspected of genital tuberculosis were included in the study. The specimen were subjected to Ziehl Neelson staining, culture on Lowenstein-Jensen media and nucleic acid amplification by Xpert MTB/RIF assay.

Results: Infertility was the most common presenting complaint in 59.77% of cases. None of the samples included in the study was positive by Ziehl – Neelson staining. Culture was positive in 4.6% of the cases. Xpert MTB/RIF assay was positive in 8.05% of cases. Compared with microscopy and culture, Xpert MTB/ RIF assay demonstrated improvement rate in detection of FGTB by 55.55%.

Conclusion: Xpert MTB/RIF assay is a useful tool for the early detection of female genital TB with the advantage of rapid results, minimal hand on time and detection of drug resistance.

 

Open Access Original Research Article

Prevalence of NS3 Mutations Inducing Resistance to Protease Inhibitors in Chronically Infected Hepatitis C Virus Genotype 4 Patients in Egypt

Ahmed Gaballah, Iman Naga, Amel Elsheredy, Gamal Elsawaf, Ola Kader

Microbiology Research Journal International, Page 1-9
DOI: 10.9734/MRJI/2018/40180

Background and Objectives: Directly acting antiviral agents, like NS3 protease inhibitors, are representing the backbone for the current therapy of Hepatitis C virus infection.

Given the heterogeneities in HCV genome, not all protease inhibitors will be equally effective against all HCV genotypes and subtypes. Data on HCV-4 mainly Egyptian strains are still inadequate.

Here we investigated the prevalence of natural NS3 mutations and Resistance-associated substitutions (RAS) among HCV-4 Egyptian isolates.

Methods: The NS5b and NS3 domains from 40 HCV Egyptian patients (20 therapy-naïve and 20 pegylated interferon-experienced patients, who failed to achieve sustained viral response) were sequenced.

Results: Using NS5b nucleotide sequences, our 40 cases were genotyped as HCV-4 of which 38 (95%) were HCV-4a, and 2 (5%) were HCV-4o.

NS3 amino acid sequence analysis showed that substitutions C16T and V36L were detected in all cases while D168H/E and T54A were found in only 3 and 1 cases, respectively. No difference could be detected between naïve and experienced patients in NS3 polymorphism.

Conclusion: Our results confirm the high genetic diversity of NS3 in HCV-4 that could impair the use of some protease inhibitors to treat HCV-4 infections. The use of cocktail therapy for HCV-4 is indispensable.

 

Open Access Original Research Article

Molecular Detection of Torque Teno Virus among HIV Seropositive Patients in Khartoum, Sudan

Ibrahim A. Almoshrf, Abdel Rahim M. El Hussein, Isam M. Elkhidir, Khalid A. Enan

Microbiology Research Journal International, Page 1-6
DOI: 10.9734/MRJI/2018/38077

Background: Torque Teno Virus (TTV) is a newly discovered non- enveloped, single-stranded DNA virus of high genotypic variability, frequently detected in patients with acute or chronic hepatitis with non A-G etiology.

Objective: This study was carried out to look for the presence of TTV among HIV seropositive patients in Khartoum State, Sudan using polymerase chain reaction (PCR) technique.

Methods: A total of 44 blood samples from HIV positive patients were tested for the presence of TTV DNA by polymer as chain reaction (PCR) using primers from UN translated (UTR) region.

Results: During the study period, 44 HIV positive patients (16male and 28female) were enrolled. Out of these TTV viri was detected in 10(22.7%) HIV positive samples.

Conclusion: The rate of TTV infection among Sudanese HIV patients was (22 %( 10/44).

 

Open Access Original Research Article

Detection of TEM and SHV Genes in Clinical Escherichia coli and Klebsiella pneumoniae Strains ESBL Isolated in Neonatology and Pediatric Units

V. M’bengue Gbonon, S. A. Afran, K. N. Guessennd, A. A. Toty, T. F. B. Diplo, A. S. P. N’Guetta, M. Dosso

Microbiology Research Journal International, Page 1-7
DOI: 10.9734/MRJI/2018/39354

Aims: To detect blaTEM and blaSHV genes in Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) strains resistant to β-lactams isolated from neonatal and pediatric infections.

Place and Duration: Bacteriology-Virology Department and Molecular Biology platform of Pasteur Institute of Côte d'Ivoire from January 2012 to November 2015.

Methods: A total of 38 strains of E. coli and K. pneumoniae ESBL isolated and identified according to classical bacteriology techniques from neonatal and pediatric infections, were subject of our study. Search for blaTEM, and blaSHV genes were carried out by conventional PCR.

Results: Molecular research showed that 52.6% of strains were carrying blaTEM gene and blaSHV gene was present in 36.9%. blaTEM and blaSHV genes were present simultaneously in 36.9% of strains.

Conclusion: This study revealed a predominance of blaTEM genes in E. coli and K. pneumoniae ESBL strains.