Development of a multiplex polymerase chain reaction assay for detection of hepatitis C virus,hepatitis B virus,and human immunodeficiency virus 1

BACKGROUND Hepatitis C virus(HCV),hepatitis B virus(HBV),and human immunodeficiency virus 1(HIV-1)are the most epidemic blood-borne viruses,posing threats to human health and causing economic losses to nations for combating the infection transmission.The diagnostic methodologies that depend on the detection of viral nucleic acids are much more expensive,but they are more accurate than sero-logical testing.AIM To develop a rapid,cost-effective,and accurate diagnostic multiplex polymerase chain reaction(PCR)assay for simultaneous detection of HCV,HBV,and HIV-1.METHODS The design of the proposed PCR assay targets the amplification of a short conserved region featured with a distinguishable melting profile and electro-phoretic molecular weight inside each viral genome.Therefore,this diagnostic method will be appropriate for application in both conventional(combined with electrophoresis)and real-time PCR facilities.Confirmatory in silico investigations were conducted to prove the capability of the approached PCR assay to detect variants of each virus.Then,Egyptian isolates of each virus were subjected to the wet lab examination using the given diagnostic assay.RESULTS The in silico investigations confirmed that the PCR primers can match many viral variants in a multiplex PCR assay.The wet lab experiment proved the efficiency of the assay in distinguishing each viral type through high-resolution melting analysis.Compared to related published assays,the proposed assay in the current study is more sensitive and competitive with many expensive PCR assays.CONCLUSION This study provides a simple,cost-effective,and sensitive diagnostic PCR assay facilitating the detection of the most epidemic blood-borne viruses;this makes the proposed assay promising to be substitutive for the mistakable and cheap serological-based assays.

Persea americana attenuates inflammatory response associated with hyperlipidemia in ovariectomized and irradiated rats by regulating MMP-3/TIMP-1 levels

Objective:To explore the effect of Persea americana supplementation on inflammation,oxidative stress,and lipid profiles in ovariectomized rats fed with a high-fat diet and exposed to radiation.Methods:The control group was sham operated,while groups 2-5 were ovariectomized and fed a high-fat diet.Groups 4 and 5 were exposed toγ-radiation(1 Gy/week for 5 weeks)after ovariectomy.Groups 3 and 5 were treated with 1 mL/250 g/day of Persea americana for one month.Serum levels of estrogen,alanine aminotransferase,aspartate aminotransferase,cholesterol,triglycerides and lipoproteins were measured.Additionally,hepatic oxidative stress,inflammatory and fibrogenic markers were evaluated.Results:Persea americana treatment reduced the oxidative stress markers as well as the levels of triglyceride,total cholesterol,and low-density lipoprotein cholesterol,which in turn lowered hepatic fat accumulation.Moreover,it suppressed hepatic inflammatory mediators(interleukin-6,tumor necrosis factor-α,and C-reactive protein)and downregulated pro-fibrogenic markers(transforming growth factor-βand tissue inhibitor of metalloproteinase-1).Conclusions:Persea americana provides protection against ovariectomy,and gamma radiation-mediated hepatic inflammation not only through its antioxidant,anti-inflammatory,lipid-lowering effect but also by modulating the fibrogenic markers.