Preparation and evaluation of rifampicin and co-trimoxazoleloaded nanocarrier against brucella melitensis infection

Background: Brucellosis or Malta fever is a contagious infection common between human and domestic animals. Many antibiotics are used for brucellosis treatment, but they are not efficient and put heavy burden on society. Co-trimoxazole and rifampicin are two candidates for brucellosis treatment. In this study, we aimed to enhance the efficacy of these antibiotics using designed nanoparticles

Methods: Different concentrations of cotrimoxazole and rifampicin were used for loading onto a nanostructure of synthesized monomethoxy poly[ethylene glycol]-oleate [mPEG-OA]. The solubility, cytotoxicity, and efficacy of these nano-packed antibiotics on Brucella-infected murine phagocytic cells were examined, as compared with free antibiotics. Then the release nanoparticles was increased approximately 3.5 and 1.5fold, respectively, which is considerable in comparison with free insoluble ones

Results: Despite acceptable loading percentage, the application of co-trimoxazole-loaded nanoparticle on Brucella-infected J774A.1 murine macrophage-like cells did not lead to reduction in the number of bacteria; however, the efficacy of rifampicin on Brucella-infected murine phagocytic cells enhanced

Conclusion: In the current study, the efficacy of rifampicin on reducing the number of Brucella melitensis increased by the novel synthesized nanostructure. In contrast, since co-trimoxazole efficacy did not enhance by loading onto nanoparticles, the co-trimoxazole inefficiency is most likely not due to its low penetration or insolubility, and probably there are other factors that remain to be clarified in the future investigations

Detection of sea, seb, sec, seq genes in staphylococcus aureus isolated from nasal carriers in Tehran province, Iran; by multiplex PCR

Staphylococcus[S.] aureus produces different extra-cellular protein toxins and virulence factors. One of the most important extra-cellular proteins is an enterotoxin which causes staphylococcal food poisoning [SFP] due to their enterotoxins. Different methods have been used to detect this toxin, each of which has advantages and disadvantages. DNA amplification methods, however, can show the presence of enterotoxigenic strains of S. aureus before the expression of enterotoxins on the basis of specific gene sequences. In this study, 150 S. aureus strains isolated from nasal carriers were confirmed by biochemical testing. PCR was used to amplify the staphylococcal enterotoxin A, B, C and Q genes, as well as the staphylococcal nuclease gene. Among the 150 healthy human isolates from the nasal carrier, 95 were confirmed as S. aureus. Only 58.9% of the isolates were diagnosed as sea, b, c, q positive. There were 24 [25.3%] isolates associated with the sea gene, 15.8% isolates associated with the seb gene, 9.5% of the isolates were associated with the sec gene, and 8.4% of the isolates associated with the seq gene. Of these isolates, 41% might be possessing additional se genes but they were not see [178 bp] and sed [319 bp] genes. The nuc gene, which encodes thermo nuclease, was used as a target DNA to identify S. aureus. Additionally, one of these enterotoxigenic isolates carried more than one toxin gene

Simultaneous molecular detection of Neisseria Meningitidis and Haemophilus Influenzae

Bacterial meningitis is a dangerous and sometimes fatal infection that affects the central nervous system. Because some antibiotics can prevent some types of these Bacteria and suppress them from spreading and infecting, therefore it is important to know what type of virus or bacterium is causing meningitis. Haemophilus influenzae and Neisseria meningitides are the two main pathogens causing acute bacterial meningitis. Different methods are used for the detection of H. influenzae and N. meningitidis but they are of low sensitivity, taking long time and difficult to perform . Therefore, complementary methods are necessary for more sensitive detection of these agents. In this study, a muliplex polymerase chain reation [mPCR] assay was developed for detection of H. influenzae and N. meningitidis. These strains were confirmed by biochemical methods. Two specific primer pairs were designed for lic-1 and opa genes of H. influenzae and N. meningitidis respectively. DNA amplification product fragments were 150 bp and 320 bp for H. influenzae and N. meningitidis, respectively. Streptococcus pneumoniae used as a negative control and did not yield a PCR product. The result of this study indicated that PCR is a useful complementary diagnostic technique, especially when Gram stain, culture, or antigenic detection is negative or inconclusive

Detection of types 40 and 41 adenoviruses in stool samples of diarrheal children by solid phase PCR

Adenoviruses [AdVs] types 40 and 41 are the causative agents of diarrhea in children. Hence, rapid sensitive and specific detection of these viruses are of clinical importance. The customary methods such as propagation of virus in cell culture suffer from limitations. Detection of immobilized amplified products in a one phase system [DIAPOPS] method has the potential to overcome these problems. A DIAPOPS method for detection of AdV types 40 and 41 was designed. Forward primers were covalently linked to the Nucleolink surface. After amplification of a 745 bp sequence of DNA binding protein gene, the amplified product was hybridized with the biotinylated probe. The hybrids were detected by the antibody-peroxidase conjugate. After optimization of the DIAPOPS conditions, 80 stool samples from children with clinical manifestation of viral diarrhea were tested. Their DIAPOPS results were compared with those of the conventional polymerase chain reaction [PCR] assay. Positive results were obtained in 11 samples. The comparison between conventional PCR and DIAPOPS showed a significant increase in sensitivity of the DIAPOPS test, 6 samples shown to be negative by conventional PCR, were demonstrated positive by DIAPOPS [p=0.00]. The DIAPOPS assay presented in this study can provide a rapid, sensitive, specific and economic method for detection of viral infections. The assay can be performed for numerous samples simultaneously in a day. This DIAPOPS method can provide a practical and reliable tool for diagnosis of enteric adenoviruses. In addition, the risk of contamination in this assay is low