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 serological 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 electrophoretic 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.

Histologic Evaluations of Xenotransplanted Rabbit Knees by In Vitro-Propagated Human Amniotic Epithelial Cells: A Preclinical Study.

OBJECTIVES: Human amniotic epithelial cells have multipotent differentiation capacity and are considered as potential therapeutic cells for clinical use. This study represents the first published report on the evaluation of the safety and clinical feasibility of human amniotic epithelial cells for transplant into knee joints, serving as an initial step for subsequent therapeutic evaluations within arthritis clinics. MATERIALS AND METHODS: Our experimental design was based on subjecting groups of rabbits as a recipient model for human amniotic epithelial cell transplant into knee joints. Twenty rabbits received 200 µL sterile 0.9% sodium chloride solution containing 1 × 109 human amniotic epithelial cells/knee joint by intra-articular injection. Control groups received cell-free saline into knees, and some animals were not treated. After 10 days of xenotransplant, radiology scans and histologic sections of transplanted and nontrans planted knees were examined and compared. Immunohistochemistry staining was also applied to detect tumor necrosis factor-alpha and interleukin 17 (as inflammatory and immuno-rejection markers) in knee sections. RESULTS: Similar to results shown in noninjected and saline-injected knees, all treated knees appeared normal, with no signs of acute immunorejection, no microbial colonization, no pain, no allergic reactions, no inflammation, and normal motion. Use of human amniotic epithelial cells appeared safe without risk of immunorejection or tumor formation in the transplanted knee joint. CONCLUSIONS: Human amniotic epithelial cells can be safely transplanted into knee joints, encouraging a need for complementary research for further therapeutic evaluations of human amniotic epithelial cells for curing arthritis.

Comparative study between topically applied irradiated human amniotic membrane in combination with tea tree oil versus topical tioconazole in pityraisis versicolor treatment.

Pityriasis versicolor (PV) is a chronic skin disease caused by virulence activities of Malassezia, a genus of skin-associated yeasts. Traditionally, Tioconazole is used as a topical antifungal for curing PV. Previous investigations cited that human amniotic membrane (HAM), a placental tissue, has antimicrobial and anti-inflammatory activities and is useful as a dressing for healing skin lesions. Moreover, tea tree oil (TTO) has a potent antifungal efficacy. This clinical trial aims to achieve an alternative therapeutic treatment able to kill Malassezia and heal PV lesions using TTO-saturated HAM (TOSHAM), with little application times. This study subjected 120 patients with hypopigmented or hyperpigmented PV lesions; half patients were treated weekly with TOSHAM compared with the others who applying 1% Tioconazole cream daily as a traditional treatment. Microbiological evaluation of in vitro fungicidal activity of TOSHAM versus Tioconazole was carried out against Malassezia furfur culture. The clinical outcomes of this study proved the superior activity of TOSHAM to heal PV lesions than Tioconazole; this was in harmony with microbiological findings. This study approached a novel therapeutic treatment of PV with great outcomes by using TOSHAM.