Optimizing Real-Time PCR methods for detection of ssaN gene Salmonella enterica subsp.enterica in the blood specimen

Background Typhoid fever caused by Salmonella typhi is a common acute infection of the reticuloendothelial system, intestinal lymphoid tissue, and gall bladder. Detection of Salmonella spp. is still based on cultures and serological methods.Widal test is one of the serological tests that is still widely used, especially in developing countries including Indonesia.Widal tests have low sensitivity and specificity. They often produce false positive or false negative results.ObjectiveThe aim of this study were i) real time PCR optimization to develop a Salmonella enterica detection system. ii) molecular detection of new target gene (ssaN gene) from blood specimens in typhoid fever patients.Methods An experimental laboratory study was performed from March to October 2016. Extraction of Salmonella typhi DNA is used as templates for the optimization of real time PCR reaction.The blood sample was from patients suspected with typhoid fever obtained from the Menteng Sub-district Health Center according to the inclusion criteria.ResultsSpecificity test of real time PCR showed that the primers and probes used are not cross-react against other microorganisms. Sensitivity test obtained minimal detection is at least 10 cfu/ml of blood specimen. In blood clinical specimens, real time PCR could detect 19 (38%) positive samples of 50 blood specimen from suspected typhoid fever patients. Eleven samples with negative Widal serology gives positive results in real time PCR.ConclusionReal time PCR used in this study can increase the level of rate of positive testing by 22% of the total specimens.Keywords : Salmonella enterica subsp.enterica, typhoid fever, ssaN gene, real time PC

Nicotiana tabacum var. Virginia Bio Oil-based Pyrolysis Extraction Have Prominence Antimicrobial Potential Compared to Ethanol Heat Reflux Extraction (EHRE)

Tobacco leaf contains antibacterial secondary metabolite compounds, such as phenol, alkaloids, and essential oils. This study compares the potential antibacterial effects of Indonesian tobacco leaf extracted using the heat reflux method (producing an extract) and pyrolysis method (providing a bio-oil). The tobacco leaf extract was challenged against Staphylococcus aureus ATCC 25923, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853. The bio-oil from the pyrolysis method showed inhibitory Kirby Bauer zones higher than those of the extract from heat reflux method, with the maximum results in the pyrolysis method indicating zones of 6.35 mm (S. aureus), 5.90 mm (E. faecalis), 3.97 mm (E. coli), and 5.025 mm (P. aeruginosa). Further study analyzed the effectiveness of the disc and well diffusion antibacterial test methods for measuring the antibacterial effect of bio-oils against Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853. The bio-oil used in the well diffusion test method showed the most significant antibacterial effectiveness. It showed the biggest inhibition zone, with a maximum of 11.65 mm and 8.90 mm for E. coli and P. aeruginosa. Our results showed Nicotiana tabacum var. Virginia Bio Oil from Ponorogo (Indonesia) is a strong potential antimicrobial, especially using well diffusion test

ANTIMICROBIAL EFFECTIVENESS OF APPLE CIDER VINEGAR IN THE GROWTH OF Staphylococcus epidermidis and Propionibacterium acnes

Acne is a skin disease caused by Propionibacterium acnes, Staphylococcus epidermidis-, and Staphylococcus aureus-induced inflammation. Apple cider vinegar contains antibacterial compounds such as acetic acid, chlorogenic acid, polyphenols, alkaloids, flavonoids, and glycosides. This study is conducted to determine the effectiveness of antibacterial compounds contained in apple cider vinegar against the growth of S.epidermidis and P.acnes, in vitro. The study used the disc diffusion method by utilizing the Mueller Hinton Agar medium. The apple cider vinegar inhibition zone was formed at the concentration of 25%, 50%, 75%, and 100% against the growth of P.acnes, whereas the inhibition zone against S.epidermidis was formed at the concentration of 50%, 75%, and 100%. The highest inhibition zone against P. acnes was 8,825 mm and against S.epidermidis was 3,725 mm at the concentration of 75%. Test results One-way ANOVA on P.acnes and Kruskal Wallis to S.epidermidis obtained (p) <0,005. This study concludes the effectiveness of apple cider vinegar in inhibiting the growth of P. acnes and S. epidermidis, where the effectiveness against P. acnes was stronger than against S. epidermidis