Coronaviruses cause disease in animals and people around the world. Human coronaviruses (HCoV) are mainly known to cause infections of the upper and lower respiratory tract but the symptoms may also involve the nervous and digestive systems. Since the beginning of December 2019, there has been an epidemic of SARS-CoV-2, which was originally referred to as 2019-nCoV. The most common symptoms are fever and cough, fatigue, sputum production, dyspnea, myalgia, arthralgia or sore throat, headache, nausea, vomiting or diarrhea (30%). The best prevention is to avoid exposure. In addition, contact per-sons should be subjected to mandatory quarantine. COVID-19 patients should be treated in specialist centers. A significant number of patients with pneumonia require passive oxygen therapy. Non-invasive ventilation and high-flow nasal oxygen therapy can be applied in mild and moderate non-hypercapnia cases. A lung-saving ventilation strategy must be implemented in acute respiratory distress syndrome and mechanically ventilated patients. Extracorporeal membrane oxygenation is a highly specialized method, available only in selected centers and not applicable to a significant number of cases. Specific pharmacological treatment for COVID-19 is not currently available. Modern medicine is gearing up to fight the new coronavirus pandemic. The key is a holistic approach to the patient including, primar-ily, the use of personal protective equipment to reduce the risk of further virus transmission, as well as patient management, which consists in both quarantine and, in the absence of specific pharmacological therapy, symptomatic treatment.

/pdf/covid-19-challenge-for-modern-medicine-4kms3i8zl8.pdf

COVID-19 challenge for modern medicine.

1 Институт биохимии и генетики – обособленное структурное подразделение Федерального государственного бюджетного научного учреждения Уфимского федерального исследовательского центра Российской академии наук, Россия, 450054, Уфа, Проспект Октября 71, E-mail: chemeris@anrb.ru 2 Башкирский государственный медицинский университет, Россия, 450008, Уфа, ул. Ленина, 3 3 ООО «Синтол», Россия, 127550, Москва, ул. Тимирязевская, д. 42 4 Институт аналитического приборостроения Российской академии наук, Россия, 198095, Санкт-Петербург, ул. Ивана Черных, д. 31/33 5 Научно-исследовательский центр Федерального государственного бюджетного учреждения «48 Центральный научно-исследовательский институт» Министерства обороны Российской Федерации, Россия, 610017, г. Киров, Октябрьский проспект, 119

Human betacoronaviruses and their highly sensitive detection by PCR and other amplification methods

This study was conducted for diagnosis and description of the pathological changes of AIV-H5 as the causative pathogen in Iraqi broiler farms. The current study was carried out on 84 broiler farms. Infected birds were tested for detection of the AIV infection from the tracheal swabs by rapid chromatographic AIV type A and H5 test kits. In RRT-PCR 8 samples (8 farms) of Trachea were selected to be tested by this assay. Samples of trachea, lung, and spleen from the dead birds with natural AIV-H5 infection were submitted for histopathological examination. seventy-two out of 84 farms tested for AIV-Type A gave positive results, and 58 out of 72 positives for type A-AIV gave a positive result for H5 antigen in a rapid chromatographic strip. The main gross lesions in the trachea of infected birds were severe congestion and hemorrhage. In the RRT-PCR assay, 8 out of 8 samples gave a distinct positive result for this test. The microscopic histopathological examination of infected tracheas showed obvious desquamation of lining epithelium with complete loss of cilia associated with congestion of blood vessels in lamina properia. Infected lungs revealed diffuse alveolar damage and severe multifocal vascular congestion. There was deposition of fibrinous material in the splenic tissue associated with the disappearance of the germinal centers. Thus, we concluded that AIV-H5 infection causes severe pathological and histopathological changes as a result of systemic infection. The RRT-PCR assay was highly sensitive and specific for the detection of highly pathogenic avian influenza virus subtypes.

/pdf/diagnosis-and-histopathological-study-of-avian-influenza-cjrq6i437n.pdf

Diagnosis and histopathological study of avian influenza virus-H5 (AIV-H5) in broiler farms

The recent recurrence of highly pathogenic avian influenza virus A H5N1 was firstly reported in mid-December 2003 and continued through 2005 This study describes a sensitive and specific real-time RT-PCR method for the detection of influenza A subtype H5 and for monitoring virus loads Using serial dilutions of influenza A H5N1 cultures, this assay reproducibly determined the lowest detection limit to be approximately 5×10−2 50 % egg infective doses (EID50) In contrast, the minimum detection limit was approximately 3 EID50 in conventional RT-PCR with WHO primers and 10 EID50 in antigen-capture ELISA In tests of serial dilutions of in vitro-transcribed influenza A H5 gene RNA, there was linear amplification from 40 copies to 4×108 copies of target RNA per reaction and approximately six copies, and sometimes even as few as three copies, of target RNA tested positive in our assay Thirty-five throat swabs from ill birds were tested: 33 samples tested positive using this assay In comparison, 27, 13 and 19 samples tested positive using conventional RT-PCR, antigen-capture ELISA and virus isolation, respectively To evaluate further the sensitivity of this real-time RT-PCR, a standard panel and 60 H5N1 isolates that contained different clades of influenza virus A/H5N1 were tested and all tested positive To evaluate the specificity of the assay, 60 throat swabs from patients infected with influenza virus A H1 were tested; all were negative Thirteen other viruses were also tested and all tested negative

Real-time RT-PCR for H5N1 avian influenza A virus detection

In the absence of effective drugs, controlling SARS relies on the rapid identification of cases and appropriate management of the close contacts, or effective vaccines for SARS. Therefore, developing specific and sensitive laboratory tests for SARS as well as effective vaccines are necessary for national authorities. Genes encoding truncated nucleocapsid (N) and spike (S) proteins of SARSCoV were cloned into the expression vector pQE30 and fusionally expressed in Escherichia coli M15. The fusion protein was analyzed for reactivity with SARS patients' sera and with anti-sera against the two human coronaviruses HCoV 229E and HCoV OC43 by ELISA, IFA and immunoblot assays. Furthermore, to evaluate the antigen-specific humoral antibody and T-cell responses in mice, the fusion protein was injected into 6-week-old BALB/c mice and a neutralization test as well as a T-cell analysis was performed. To evaluate the antiviral efficacy of immunization, BALB/c mice were challenged intranasally with SARSCoV at day 33 post injection and viral loads were determined by fluorescent quantitative RT-PCR. Serological results showed that the diagnostic sensitivity and specificity of the truncated S-N fusion protein derived the SARS virus were > 99% (457/460) and 100.00% (650/650), respectively. Furthermore there was no cross-reactivity with other two human coronaviruses. High titers of antibodies to SRASCoV appeared in the immunized mice and the neutralization test showed that antibodies to the fusion protein could inhibit SARSCoV. The T cell proliferation showed that the fusion protein could induce an antigen-specific T-cell response. Fluorescent quantitative RT-PCR showed that BALB/c mice challenged intranasally with SARSCoV at day 33 post injection were completely protected from virus replication. The truncated S-N fusion protein is a suitable immunodiagnostic antigen and vaccine candidate.

/pdf/the-expression-and-antigenicity-of-a-truncated-spike-srsofn6p40.pdf

The expression and antigenicity of a truncated spike-nucleocapsid fusion protein of severe acute respiratory syndrome-associated coronavirus.

https://link.springer.com/content/pdf/10.1007%2FBF03260067.pdf

Real-time quantitative fluorescent reverse transcriptase-PCR for detection of severe acute respiratory syndrome-associated coronavirus RNA.

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Real-time quantitative fluorescent reverse transcriptase-PCR for detection of severe acute respiratory syndrome-associated coronavirus RNA.