A highly efficient and accurate method of detecting and subtyping Influenza A pdm H1N1 and H3N2 viruses with newly emerging mutations in the matrix gene in Eastern Taiwan.

The rapid identification of Influenza A virus and its variants, which cause severe respiratory diseases, is imperative to providing timely treatment and improving patient outcomes. Conventionally, two separate assays (total test duration of up to 6 h) are required to initially differentiate Influenza A and B viruses and subsequently distinguish the pdm H1N1 and H3N2 serotypes of Influenza A virus. In this study, we developed a multiplex real-time RT-PCR method for simultaneously detecting Influenza A and B viruses and subtyping Influenza A virus, with a substantially reduced test duration. Clinical specimens from hospitalized patients and outpatients with influenza-like symptoms in Eastern Taiwan were collected between 2011 and 2015, transported to Hualien Tzu Chi Hospital, and analyzed. Conventional RT-PCR was used to subtype the isolated Influenza A viruses. Thereafter, for rapid identification, the multiplex real-time RT-PCR method was developed and applied to identify the conserved regions that aligned with the available primers and probes. Accordingly, a multiplex RT-PCR assay with three groups of primers and probes (MAF and MAR primers and MA probe; InfAF and InfAR primers and InfA probe; and MBF and MBR primers and MB probe) was established to distinguish these viruses in the same reaction. Thus, with this multiplex RT-PCR assay, Influenza B, Influenza A pdm H1N1, and Influenza A H3N2 viruses were accurately detected and differentiated within only 2.5 h. This multiplex RT-PCR assay showed similar analytical sensitivity to the conventional singleplex assay. Further, the phylogenetic analyses of our samples revealed that the characteristics of these viruses were different from those reported previously using samples collected during 2012-2013. In conclusion, we developed a multiplex real-time RT-PCR method for highly efficient and accurate detection and differentiation of Influenza A and B viruses and subtyping Influenza A virus with a substantially reduced test duration for diagnosis.

Spontaneous spinal epidural hematoma: a case report.

BACKGROUND: Spinal epidural hematomas usually occur under certain conditions; they rarely occur spontaneously. The prevalence of spontaneous spinal epidural hematoma is ~ 0.1 per 100,000, and the male-to-female ratio is approximately 1.4 to 1. Herein, we describe a rare case of spontaneous spinal epidural hematoma. CASE PRESENTATION: A 63-year-old Taiwanese woman, with underlying hypertension, anemia, and a history of cardiovascular accident without sequela, was admitted to our emergency department with a chief complaint of sudden bilateral weakness in the lower limbs. Magnetic resonance imaging revealed a spontaneous epidural hematoma. The patient underwent emergency surgery to remove the epidural hematoma and laminectomy for decompression. The bilateral lower limb weakness was alleviated immediately after the surgery. CONCLUSION: In patients with no risk factors related to spinal epidural hematoma, symptoms of bilateral lower limb weakness must be investigated carefully because this condition may occur spontaneously.

Aspirin plus tirofiban inhibit the thrombosis induced by Russell's viper venom.

BACKGROUND: Thrombosis and coagulopathy are the commonest hematological manifestations of envenomation of Russell's viper venom (RVV). Factor X is activated by a factor X-activating enzyme from Russell's viper venom (RVV-X) to start the coagulation cascade. We established an animal model with local ischemic effects induced by RVV. We tried to treat RVV envenomation with antiplatelets and anticoagulants without recourse to antivenom. METHODS: RVV was injected into the foot pad of mice. We observed the effects at different intervals and compared local changes in ischemia with drug treatment after 30 min. RESULTS: A combination of aspirin plus tirofiban could prevent the ischemic change induced by RVV. The antithrombotic effects of single-use of aspirin or tirofiban were better than single-use of heparin or clopidogrel. CONCLUSION: The aspirin + tirofiban group had a better outcome with respect to prevention of tissue ischemia and gangrene. This indicates that the activation and aggregation of platelets is the major cause of thrombosis induced by RVV.