Novel Corona Virus: One Biological Disaster of 2020

 

Ghogare Rajashree D., Navale Abhijit S., Shaikh Sahil B

M. Pharm Pharmacology, Department of Pharmacology, Pravara Rural College of Pharmacy,

Pravaranagar,  Rahata, Ahmednagar.

*Corresponding Author E-mail: rajashreeghogare247@gmail.com

 

Abstract:

All the patients are reported that various respiratory diseases such as common cold as mild and pneumonia as in savior condition and other symptoms are also seen in some cases such as Radiographic ground lung changes, lower white blood cell lymphocytes and platelet count, hypoxemia and deranged liver and renal function etc has formed. The novel coronavirus (2019-nCoV) was first found from a patient with pneumonia which is linked to the cluster of acute respiratory illness cases from Wuhan, China. The cells were observed daily for cytopathic effects by light microscopy and the cell supernatants were collected for use in quantitative RT-PCR assays. Coronaviruses cause a large variety of diseases in some animals, and their ability to cause severe disease in livestock and companion animals such as pigs, cows, chickens, dogs and cats led to significant research on these viruses in the last few year of the 20th century. Unexplained pneumonia, Radiographic ground glass lung changes, Lowering of white blood cell lymphocyte, Lowering of white blood cell pletlet, Hypoxemia, Derangd liver and Renal function etc are conclude savior symptoms of this disease. Coronavirus, Seafood, Mouse hepatitis virus, these are the causative agent of this disease. In January 2020, several organizations and institutions began work on creating Vaccine research vaccines for 2019 n-CoV based on the published genome.

 

KEYWORDS: Novel Coronavirus 2019, unexplained pneumonia, PCR technique, SARS- Severe Acute Respiratory Syndrome, MERS-Middle East Respiratory Syndrome.

 

 


INTRODUCTION:

Novel coronavirus is major group of coronavirus; this virus is mainly found in wuhan city of china, originated from sea food market. The code of genetic material of this virus is surrounded by enveloped with protein spikes which looks as Crown therefore named as CORONA. Coronavirus is a RNA virus. It is positive sense RNA virus, with linear structure. This virus is single stranded and the capsids of virus are helices.

 

This virus is nonliving virus because it lacks cellular structure but it replicates very quickly. First case of novel coronavirus were reported on Dec 31, 2019 but within a month 20,629 cases were reported from which 427 has dead in China up to Feb 3, 2020.Now days it will also spread in India by the travelers of China to India. Initially 9 cases were reported in INDIA from which 2 are from Kerala, 3 from Pune, 1 from Delhi, and 3 from Mumbai. All the patients who were reported are common for various respiratory diseases such as common cold as mild and pneumonia as savior condition. The other symptoms are also seen in some cases such as Radiographic ground lung changes, lower white blood cell lymphocytes and platelet count, hypoxemia and deranged liver and renal function.

 

Historical background:

On 31 December 2019, the Wuhan Municipal Health Commission in Wuhan City, Hubei province, China reported a cluster of 27 pneumonia cases of aetiology is unknown, In that seven severe cases reported link to Wuhan's Huanan that Seafood Wholesale Market. The cases presented with clinical symptoms are common to several infectious respiratory diseases like fever, dyspnoea, and bilateral lung infiltrates on chest radiographs. The market was closed for public from 1 January 2020 as per authorities all cases are under observation.

 

As per report of China CDC on 9 January 2020, the novel coronavirus (2019-nCoV) had been detected as the causative agent for 15 of the 59 pneumonia cases. On 10 January 2020, the first novel coronavirus genome sequence was ready to public availability then the sequence was deposited in the Gen Bank database and was uploaded to the Global Initiative on Sharing all Influenza Data (GISAID). Preliminary analysis showed that the novel coronavirus (2019-nCoV) clusters with the SARS-related CoV clade and differs from the core genome of known bat CoVs.

 

It is not clear to ECDC if cases have been hospitalised only due to medical needs, or also for isolation purposes for milder cases. In Guangdong, from reported 14 cases, In that two had not travelled to Wuhan, China, but had a history of contact with confirmed cases. The other four laboratory-confirmed cases are travelling similarly two reported from Thailand, one from Japan, while one from South Korea.

 

The four reported deaths, all in China, the first death occurred on 9 January 2020 in a 61-year-old patient with severe underlying conditions, who reportedly visited the Wuhan’s Huanan Seafood Wholesale market all year round. A second death occurred in a 69-year-old case on 15 January 2020 with multiple organ failure. The third death was reported on 18 January 2020, and the fourth death occurred in an 89-year-old with pre-existing medical conditions on 19 January 2020. In China, 1739 close contacts have been identified and monitored. Of these, 817 have completed the observation period, while 922 remain under medical observation [6]

 

Fig no1: Morphological structure of virus

 

Genetic sequence of corona virus:

Genetic sequencing of the Novel coronavirus offers leads to its initiate and spread. More than 20 strains of infected volunteers have been sequenced in the laboratories of China and Thialand and have made them freely assessable to public. Bedford trevor who is evolutionary geneticist from fred Hutchinson cancer search centre in seattle and other geneticists are using the free assessable data of infected volunterrs to determine when the virus emerged current evaluate point to November 2019. Viral sequences, Bedford adds that, he could identify any genetic changes that might have helped the virus to make them acquired from animals to humans. And if there is extensive human-to-human transmission, Bedford and other geneticists will be looking for signs that the virus has increase further mutations that are enabling it to spread more efficiently in humans. Bedford cautions that any conclusions are basic, because very less data is available.(3)

 

Fig no 2: Structure of corona virus

Types of Corona virus:

1.     229E and OC43:

(unranked): Virus, Realm: Riboviria, Phylum: incertae sedis, Order: Nidovirales, Family: Coronaviridae, Genus: Alphacoronavirus, Subgenus: Duvinacovirus, Species: Human coronavirus 229E HCoV-229E is associated with large number of clinical respiratory manifestation, including the common cold to high-morbidity outcomes like pneumonia and bronchiolitis. It is also frequently analogues with other respiratory viruses, particularly with human respiratory syncytial virus (HRSV). HCoV-229E is one of the seven human coronaviruses which involved HCoV-NL63, HCoV-OC43, and HCoV-HKU1 which are globally spraded. However, the viruses were detected in different parts of the world in different years.(10)

 

2.     SARS and MERS:

Coronaviruses were found in the mid-1960s which are known to infect humans and a variety of animals (including birds and mammals). The primary goal of cells are epithelial cells in respiratory and G.I tract. Transmission can be done by a various routes such as respiratory droplets, airborne, fomites or faecal-oral. Till now seven coronaviruses have been shown to infect humans. Common human coronaviruses Betacorona virus HCoV-OC43 and HCoV-HKU1, as well as Alphacoronavirus HCoV-229E can cause common colds and also severe lower respiratory tract infections, pneumonia in the youngest and oldest age groups. Since 2002, two additional coronaviruses infecting animals have detected and caused eruption in humans: SARS-CoV (2002, Betacoronavirus, subgenus Sarbecovirus), and MERS-CoV (2012, Betacoronavirus, subgenus Merbecovirus).(10)

 

3.     HCoV NL 63(Human Corona Virus NL 63):

(unranked): Virus Realm: Riboviria, Phylum: incertae sedis, Order: Nidovirales, Family: Coronaviridae, Genus: Alphacoronavirus, Subgenus: Setracovirus, Species: Transmission of HCoV-NL63 is occur through droplet expulsion from the respiratory tract, which may be airborne or spread through close personal contact. At room temperature the virus is able to survive for seven days in respiratory secretions and remains infective. After entering into host the virus has binds to cellular receptors using spike proteins, same as that of found in HIV-1.The virus is able to use Angiotensin-converting enzyme 2 (ACE2) as an entry receptor to bind to and enter target cell. The specific entry of the virus into the host cell has been completed is not determined yet. Therefore, entrance] into the cell is either through direct cell fusion with the plasma membrane or endocytosis. (12)

 

4.     HKU-1:

(Unranked): Virus Realm: Riboviria ,Phylum: Incertae Sedis , Order: Nidovirales, Family: Coronaviridae Genus: Betacoronavirus, Subgenus: Embecovirus, Species: Human coronavirus HKU1 HCoV-HKU1 was first recognized in January, 2005, in a 71-year-old man who was hospitalized with an acute respiratory distress and radiology graphically confirmed unexplained pneumonia. He had recently returned to Hong Kong from Shenzhen, China.[11]

 

5.     Novel coronavirus (2019-nCoV):

The novel coronavirus (2019-nCoV) was first detected from a patient with pneumonia which is linked to the cluster of acute respiratory illness cases from Wuhan, China. Genetic analysis confirmed that it is closely related to SARS-CoV and genetically clusters within the genus: Betacoronavirus, subgenus: Sarbecovirus. There is currently limited information on the epidemiological and clinical features of the infection caused by 2019-nCoV. Case definitions for the European Region are currently under review by ECDC and WHO Regional Office for Europe.(6)

 

Virus isolation:

For virus isolation special-pathogen-free human airway epithelial (HAE) cells were used. In short, bronchoalveolar lavage fluids or throat swabs from the volunteers were vaccinate into the HAE cells through the apical surfaces HAE cells were maintained in an air–liquid interface incubated at 37°C. The cells were observed daily for cytopathic effects by light microscopy and the cell supernatants were collected for use in quantitative RT-PCR assays. Apical samples were collected for sequencing after three samples(5)

 

Coronavirus Life Cycle:

 

Fig no.2: Life cycle of corona virus of different species

 

1.     Attachment and Entry:

The primary joining of the virion to the host cell is introduced by interactions between the S protein and its receptor. The sites of receptor binding domains (RBD) in the S1 region of a coronavirus S protein changes depend on the virus, with some having the RBD at the N-terminus of S1 (MHV) while others (SARS-CoV) have the RBD on the C-terminus of S1. The S-protein/receptor interaction is the initial determinant for a coronavirus to infect a host species and also governs the tissue tropism of the virus. Peptidases are useful to coronavirus as their cellular receptor. It is not clear why peptidases are used, as entry takes place even in the absence of the enzymatic domain of such proteins. Many α-coronaviruses use aminopeptidase N (APN) as their receptor, SARS-CoV and HCoV-NL63 utilize angiotensin-converting enzyme 2 (ACE2) as their receptor, MHV enters through CEACAM1, and the recently identified MERS-CoV binds to dipeptidyl-peptidase 4 (DPP4) to get entre into human cells. The virus must then gain access to the host cell cytosol. This is generally skilled by acid-dependent proteolytic breaking of S protein by a cathepsin, TMPRRS2 or other protease is followed by the fusion of the viral and cellular membranes. S protein breaking occurs at two sites within the S2 portion of the protein, with the first cleavage important for separating the RBD and fusion domains of the S protein and the second for uncovering the fusion peptide (cleavage at S2′). Fusion generally happen within acidified endosomes, but some coronaviruses, like MHV, can fuse at the plasma membrane. Cleavage at S2′ exposes a fusion peptide that slot in the membrane, which is followed by joining of two heptad repeats in S2 forming an antiparallel six-helix bundle. This formation of bundle allows for the mixing of viral and cellular membranes which results in fusion and ultimately release of the viral genome in the cytoplasm. (14)

 

2.     Replicase Protein Expression:

The further step in the coronavirus lifecycle is the relocation of the replicase gene from the virion genomic RNA. The replicase gene encodes two large ORFS, rep1a and rep1b, which express two co-terminal polyproteins, pp1a and pp1ab. For expressing both polyproteins, the virus uses a slippery sequence (5′-UUUAAAC-3′) and an RNA pseudo knot which results in ribosomal frame shifting from the rep1a reading frame into the rep1b ORF. In most cases, the ribosome uncoils the pseudo knot structure, and continues translation until it encounters the rep1a stop codon. Sometimes the pseudo knot blocks the ribosome from continuing elongation, causing it to pause on the slippery sequence, which changes the reading frame by moving back one nucleotide, −1 frame shift, before the ribosome is able to melt the pseudo knot structure and extend translation into rep1b, results in the translation of pp1ab.

 

In vitro studies predict the incidence of ribosomal frame shifting to be as high as 25%, but this has not been discovered in the context of virus infection. It is unknown that exactly why these viruses utilize frame shifting to control protein expression, but it is hypothesized to either control the precise ratio of rep1b:rep1a proteins or delay the production of rep1b products until the products of rep1a have prepared an acceptable environment for RNA replication.

 

Polyproteins pp1a and pp1ab have the nsps 1–11 and 1–16, respectively. In pp1ab, nsp11 from pp1a becomes nsp12 following extension of pp1a into pp1b.where γ-coronaviruses do not contain a comparable nsp1. These polyproteins are subsequently cleaved into the individual nsps. Coronaviruses may encode either two or three proteases that cleave the replicase polyproteins. They are the papain-like proteases (PLpro), encoded within nsp3, and a serine type protease, the main protease, or Mpro, encoded by nsp5. Most coronaviruses encode 2 PLpros within nsp3, except the γ-coronaviruses, SARS-CoV and MERS-CoV which can express only one PLpro.

 

The PLpros cleave the nsp1/2, nsp2/3, and nsp3/4 boundaries while the Mpro is responsible for the remaining 11 cleavage events. Further, many of the nsps assemble into the replicase-transcriptase complex (RTC) to form an environment suitable for RNA synthesis, and ultimately are responsible for RNA replication and transcription of the sub-genomic RNAs. The nsps also contain other enzyme domains and functions, including those important for RNA replication, for example nsp12 encodes the RNA-dependent RNA polymerase (RdRp) domain; nsp13 encodes the RNA helicase domain and RNA 5′-triphosphatase activity; nsp14 encodes the exoribonuclease (ExoN) involved in replication fidelity and N7-methyltransferase activity; and nsp16 encodes 2′-O-methyltransferase activity.

 

In addition to the replication functions other activities, like blocking innate immune responses (nsp1; nsp16-2′-O-methyl transferase; nsp3-deubiquitinase) have been recognized for some of the nsps, with other large nuber of unknown functions (nsp3-ADP-ribose-1”-phosphatase; nsp15-endoribonuclease (NendoU)) also identified. For a list of non-structural proteins and their proposed functions, Interestingly, ribonucleases nsp15-NendoU and nsp14-ExoN activities are different to the Nidovirales order and are considered genetic markers for such viruses. (13)

 

3.     Replication and Transcription:

Viral RNA synthesis follows the translation and assembly of the viral replicase complexes. Both genomic and sub-genomic RNAs are produce by such complexes. Sub-genomic RNAs serve as mRNAs for the structural and extra genes which reside downstream of the replicase polyproteins. All positive-sense sub-genomic RNAs are 3′ co-terminal with the full-length viral genome and it forms a set of nested RNAs, a distinctive property of the order Nidovirales. Both genomic and sub-genomic RNAs are obtained through negative-strand intermediates. These negative-strand intermediates are only about 1% as abundant as their positive-sense counterparts and have both poly-uridylate and anti-leader sequences.

 

Most cis-acting sequences are important for the replication of viral RNAs. In the 5′ UTR of the genome are seven stem-loop structures that may extend into the replicase 1a gene. The 3′ UTR contains a bulged stem-loop, a pseudoknot, and a hypervariable region. Interestingly, the stem-loop and the pseudoknot at the 3′ end overlap, and thus cannot form simultaneously. Therefore, such different structures are proposed to regulate alternate stages of RNA synthesis, although exactly which stages are regulated and their precise mechanism of action are not kown yet.

 

Perhaps the most new aspect of coronavirus replication is how the leader and body TRS segments fuse during production of sub-genomic RNAs. This was originally thought to take place during positive-strand synthesis, but now it is mostly believed to occur during the discontinuous extension of negative-strand RNA. The current model introduces that the RdRp pauses at any one of the body TRS sequences (TRS-B); following this pause the RdRp either continues elongation to the next TRS or it switches to amplifying the leader sequence at the 5′ end of the genome guided by complementarity of the TRS-B to the leader TRS (TRS-L). Many pieces of evidence currently support this model, including the presence of anti-leader sequence at the 3′ end of the negative-strand sub-genomic RNAs. However, many questions remain to fully define the model.

 

For instance, how does the RdRp bypass all of the TRS-B sequences to produce full-length negative-strand genomic RNA? Also, how are the TRS-B sequences directed to the TRS-L and how much complementarity is necessary? Answers to these questions and others will be necessary to gain a full perspective of how RNA replication occurs in coronaviruses.(13)

 

Finally, coronaviruses are also known for their ability to recombine using both homologous and non-homologous recombination. The ability of these viruses to recombine is tied to the strand switching ability of the RdRp. Recombination likely plays a prominent role in viral evolution and is the basis for targeted RNA recombination, a reverse genetics tool used to engineer viral recombinants at the 3′ end of the genome.(14)

4.     Assembly and Release

Further replication and sub genomic RNA synthesis, the viral structural proteins, S, E, and M are translated and inserted in the endoplasmic reticulum (ER). These proteins move through the secretory pathway in the endoplasmic reticulum-Golgi intermediate compartment (ERGIC). There, viral genomes encapsulated by N protein bud into membranes of the ERGIC containing viral structural proteins, forms mature virions. The M protein directs most protein-protein interactions recommended for assembly of coronaviruses. However, M protein is not sufficient for virion production, as virus-like particles (VLPs) cannot be formed by M protein expression alone. However, when M protein is expressed along with E protein VLPs are formed, suggesting these two proteins function together to produce coronavirus envelopes. N protein enhances VLP formation, suggested that fusion of encapsulated genomes in the ERGIC enhances viral envelopment.

 

The S protein is incorporated into virions at this step, but is not required for assembly. The ability of the S protein to traffic to the ERGIC and interact with the M protein is critical for its incorporation into virions. While the M protein is relatively abundant, the E protein is only present in small quantities in the virion. Thus, it is likely that M protein interactions provide the impetus for envelope maturation. It is not known how E protein assists M protein in assembly of the virion, and several possibilities have been suggested. Some work has indicated a role for the E protein in inducing membrane curvature, although others have suggested that E protein prevents the aggregation of M protein. The E protein may also have another role in promoting viral release by indicating the host secretory pathway (14)

 

The M protein also binds to the nucleocapsid, and this interaction promotes the completion of virion assembly. These interactions have been mapped to the C-terminus of the endodomain of M with CTD 3 of the N-protein. However, it is not clear exactly how the nucleocapsid complexed with virion RNA traffics to the ERGIC to interact with M protein and become incorporated into the viral envelope. Another outstanding question is how the N protein selectively packages only positive-sense full-length genomes among the many different RNA species produced during infection. (12) A packaging signal for MHV has been identified in the nsp15 coding sequence, but mutation of this signal does not appear to affect virus production, and a mechanism for how this packaging signal works has not determined yet.

 

Furthermore, number of coronaviruses do not contain similar sequences at this locus, indicates that packaging may be virus specific. In below given assembly, virions are transported to the cell surface in vesicles and released by exocytosis. It is not known if the virions use the traditional pathway for transport of large cargo from the Golgi or if the virus has diverted a separate, different pathway for its own exit. In some of the coronaviruses, S protein that does not get assembled in virions transits to the cell surface where it mediates cell-cell fusion between infected cells and adjacent, uninfected cells. This guide to the formation of giant, multinucleated cells, that allows the virus to spread within an infected organism without being recognized or neutralized by virus-specific antibodies.(13)

 

Pathogenesis:

Animal Coronaviruses:

Coronaviruses cause a large variety of diseases in animals, and their ability to cause severe disease in livestock and companion animals such as pigs, cows, chickens, dogs and cats led to significant research on these viruses in the last few year of the 20th century.

                                                                                                    

Let us assumed that, transmissible Gastroenteritis Virus (TGEV) and Porcine Epidemic Diarrhea Virus (PEDV) formed severe gastroenteritis in young piglets, which leads to significant morbidity, mortality, and ultimately it is hazardous to economic.

                                                                                                    

In North America PEDV is emersed recently for the first time, causing significant losses of young piglets. Porcine hemagglutinating encephalomyelitis virus (PHEV) mostly leads to enteric infection but has to able infect the nervous system which can cause encephalitis, vomiting and wasting in pigs.(18) Feline enteric coronavirus (FCoV) causes a mild or asymptomatic infection in domestic cats, but during persistent infection, mutation transforms the virus into a highly virulent strain of FCoV (Feline Infectious Peritonitis Virus, FIPV), that leads to development of a lethal disease knows as feline infectious peritonitis (FIP).

                                                                                                    

FIP has wet and dry forms, with similarities to the human disease, sarcoidosis. FIPV is macrophage tropic and it is believed that it causes aberrant cytokine and chemokine expression and lymphocyte depletion, resulting in lethal disease. But additional research is needed to confirm this hypothesis. Bovine CoV, Rat CoV, and infectious Bronchitis Virus (IBV) cause mild to severe respiratory tract infections (RTI) in cattle, rats, and chicken. Bovine CoV causes significant losses in the cattle industry and also has spread to infect a variety of ruminants, including elk, deer and camels(14)

                                                                                                    

In addition to severe respiratory disease, the virus causes diarrhoea, it all initiate to weight loss, dehydration, inhibition of milk production, Some strains of IBV, a γ-coronavirus, it also affect the uro-genital tract of chickens which causes renal disease. IBV significantly diminishes egg production and weight gain, causing substantial losses in the chicken industry each year. Now a day a novel coronavirus named SW1 was found in a deceased Beluga whale. (17) Large numbers of virus particles were identified in the liver of the deceased whale with respiratory disease and acute liver failure. The electron microscopic images were not sufficient to detect the virus as a coronavirus; sequencing of the liver tissue clearly identified the virus as a coronavirus.

                                                                                                    

It was subsequently determined to be a γ-coronavirus based on phylogenetic analysis but it has not yet been verified experimentally that this virus is these viruses are highly divergent from other nido viruses but are similarly related to the roniviruses. In size, they are 20 kb, falling in between large and small nidoviruses. Interestingly, these viruses do not encode for an endoribonuclease, which is present in all other nidoviruses. These attributes suggest these viruses are the prototype of a new nidovirus family and may be a missing link in the transition from small to large nidoviruses. Actually whales are a causative agent of disease. In addition, there has been intense interest in identifying novel bat CoVs, since these are the likely ultimate source for SARS-CoV and MERS-CoV, and hundreds of novel bat coronaviruses have been identified over the past decade. Finally, another novel group of nidoviruses, Mesoniviridae, were recently identified as the first nidoviruses to exclusively infect insect hosts(14)

 

Symptoms:

1.     Sevior Symptoms:

Unexplained pneumonia, Radiographic ground glass lung changes, Lowering of white blood cell lymphocyte, Lowering of white blood cell pletlet, Hypoxaemia, Derangd liver and Renal function, Respiratory symptoms and pulmonary infilrates on chest radiograph, Pneumonia, Kidney failure(11)

2.     Common symptoms:

Fever, Common cold, Sneezing, Coughing, Diarrhea, Chest pain, Generalised weakness, Shortness of breath (1)

 

Diagnosis:

A.   Molecular diagnostics for 2019-nCoV:

On the basis of the genome sequences obtained, a real-time PCR detection assay was developed. PCR primers and probes were designed using Applied biosystems Primer Express Software (Thermo Fisher Scientific, Foster City, CA, USA) on the basis of our sequenced virus genomes. The specific primers and probe set (labelled with the reporter 6-carboxyfluorescein [FAM] and the quencher Black Hole Quencher 1 [BHQ1]) for orf1a were as follows:

 

Forward primer5ʹ-AGAAGATTGGTTAGATGATGATAGT-3ʹ;

 

Reverse primer5ʹ TTCCATCTCTAATTGAGGTTGAACC-3ʹ; and

 

Probe 5ʹ-FAM-TCCTCACTGCCGTCTTGTTGACCA BHQ1-3ʹ.

 

The human GAPDH gene was used as an internal control (forward primer 5ʹ TCAAGAAGGTGGTGAAGCAGG- 3ʹ; reverse primer5ʹ-CAGCGTCAAAGGTGGAGGAGT-3ʹ; probe 5ʹVICCCTCAAGGGCATCCTGGGCTACACTBHQ1- 3ʹ).

 

Primers and probes were synthesised by BGI (Beijing, China). RT-PCR was done with an Applied Biosystems 7300 Real-Time PCR System (Thermo Scientific), with 30 μL reaction volumes consisting of 14 μL of diluted RNA, 15 μL of 2X Taqman One-Step RT-PCR Master Mix Reagents (4309169; Applied Biosystems, Thermo Fisher), 05 μL of 40X Multi Scribe and RNase inhibitor mixture, 0.75 μL forward primer (10 μmol/L), 0.75 μL reverse primer (10 μmol/L), and 0.375 μL probe (10 μmol/L). Thermal cycling parameters were 30 min at 42°C, followed by 10 min at 95°C, and a subsequent 40 cycles of amplification (95°C for 15 s and 58°C for 45 s). Fluorescence was recorded during the 58°C phase (5)

 

Phylogenetic analysis:

Sequence alignment of 2019-nCoV with reference sequences was done with Mafft software Phylogenetic analyses of the complete genome and major coding regions were done with RAxML software 22 with 1000 bootstrap replicates, employing the general time reversible nucleotide substitution model (5)

 

Etiological agent:

Coronavirus, Seafood, Mouse hepatitis virus, Murinae, Tylonycteris bat coronavirus HKU4, Rousettus bat coronavirus HKU9 (11), HCoV-229E transmits via dropletrespiration and fomite, Virulence, Human coronavirus HKU1(10)

 

Treatment:

(The treatment of corona viruses is not known the total treatment is predictable)

Sr. No

Category of Drug

1

Oxygen therapy

2

Air purifier ,Ventilation non-invasive→face mask, etc

3

Ventilation invasive

4

Continuous Renal Replacement Therapy (CRRT)

5

 Extra corporeal membrane oxygenation (ECMO)

6

Antibiotic

7

Antifungal

8

Antiviral

9

Glucocorticoids

10

Intravenous immunoglobulin

 

The disease caused by 2019-nCoV has been temporarily designated "2019-nCoV acute respiratory disease" by the WHO. No specific treatment verified by medical research standards (in the sense of systematic reviews of peer reviewed randomized controlled clinical trials) is available as of February 2020, so treatment is focused on alleviation of symptoms, which may include fever, dry cough, and shortness of breath In parallel, multiple lines of exploratory research were started into potential treatments of the disease in January 2020.

 

The Chinese Center for Disease Control and Prevention started testing existing pneumonia treatments for efficacy in treating coronavirus-related pneumonia in late January. Investigations into the effectiveness of existing antivirals, including protease inhibitors like indinavir, saquinavir and lopinavir /ritonavir also started in late January. Examination of the RNA polymerase inhibitor remdesivir, interferon beta, and previously identified monoclonal antibodies (mAbs) as possible treatments also started around the same period. Projects studying the effectiveness of Hepatitis C treatment sofosbuvir, a RNA-dependent RNA polymerase inhibitor, were also started in late January 2020.

 

Researchers from the Norwegian University of Science and Technology (NTNU) have created a database with 120 safe-in-man broad spectrum antiviral agents and identified 31 drug candidates for treatment of 2019- nCoV. In late January 2020, Chinese medical researchers expressed an intent to start clinical testing on remdesivir, chloroquine, and lopinavir /ritonavir, all of which seemed to have "fairly good inhibitory effects" on 2019-nCoV at the cellular level in exploratory research.(16) On 2 February 2020, doctors in Thailand claimed to have treated a patient successfully with a combination of lopinavir/ritonavir and the influenza drug oseltamivir. (14)

 

Vaccine research:

In January 2020, several organizations and institutions began work on creating Vaccine research vaccines for 2019 n-CoV based on the published genome. In China, the Chinese Center for Disease Control and Prevention is developing a vaccine against the novel coronavirus .The team of Yuen Kwokyung at the University of Hong Kong, which previously participated in work on the SARS coronavirus during its 2003 outbreak, has also announced that a vaccine is under development there but has yet to proceed to animal testing.

 

Shanghai East Hospital is also developing a vaccine in partnership with the biotechnology company Stemirna Therapeutics. Elsewhere, three vaccine projects are being supported by the Coalition for Epidemic Preparedness Innovations (CEPI), including projects by the biotechnology companies Moderna and Inovio Pharmaceuticals and another by the University of Queensland. The United States National Institutes of Health (NIH) is cooperating with Moderna to create an RNA vaccine matching a spike of the coronavirus surface, and is hoping to start production by May 2020.

 

Invivo Pharmaceuticals is developing a DNAbased vaccination and collaborating with a Chinese firm in order to speed its acceptance by regulatory authorities in China, hoping to perform human trials of the vaccine in the summer of 2020. In Australia, the University of Queensland is investigating the potential of a molecular clamp vaccine that would genetically modify viral proteins to make them mimic the coronavirus and stimulate an immune reaction.

 

In an independent project, the Public Health Agency of Canada has granted permission to the International Vaccine Centre (VIDO-InterVac) at the University of Saskatchewan to begin work on a vaccine.[14] VIDO-Inter Vac aims to start production and animal testing in March 2020, and human testing in 2021.The Imperial College Faculty of Medicine in London has funding to develop a vaccine and take it to animal testing, a phase of research it expects to complete by mid-February 2020.

 

CONCLUSION:

Last 50 years the emergence of many different coronaviruses that cause a various types of human and veterinary diseases has occurred. It is likely that these viruses will continue to emerge and to evolve and cause both human and veterinary outbreaks owing to their ability to recombine, mutate, and infect multiple species and cell types. Future research on coronaviruses will continue to investigate many aspects of viral replication and pathogenesis. First, understanding the propensity of these viruses to jump between species, to establish infection in a new host, and to identify significant reservoirs of coronaviruses will dramatically aid in our ability to predict when and where potential epidemics may occur. As bats seem to be a significant reservoir for these viruses, it will be interesting to determine how they seem to avoid clinically evident disease and become persistently infected. According to different literature survey the effect of the viruse are shown in for of graphical representation by help of observation shown in bellow

 

RESULT:

According to my opinion and study about virus the limit human-to-human transmission including reducing secondary infections among close contacts and health care workers, preventing transmission amplification events, and preventing further international spread from China. Identify isolate and care for patients early, including providing optimized care for infected patients. Identify and reduce transmission from the animal source Address crucial unknowns regarding clinical severity, extent of transmission and infection, treatment options, and accelerate the development of diagnostics, therapeutics and vaccines.

 

ACKNOWLEDGEMENT:

By the help of different lecture and article survey has assumed that the corona viral disease is show very dangerous effect on human as well as animal habitat with vigorously spreading. The corona virus is having different form and usually having different harmful effect but novel corona virus is showing tremendous antibiotic resistance and other drug therapy also. For that virus inhibit similarly drug therapy like generally pneumonia disease is used but not effectively work hence new vaccines are developed successfully. Now a days to control of inhibition of this viral disease is done successfully.

 

REFERENCE:

1.      Jasper Fuk-Woo Chan, Shuofeng Yuan, Kin-Hang Kok, Kelvin Kai-Wang To, Hin Chu, A familial cluster of pneumonia associated with the 2019 novel coronavirus indicating person-to-person transmission: a study of a family cluster; Published online January 24, 2020

2.      Yvonne Xinyi Lim, Yan Ling Ng, James P. Tam and Ding Xiang Liu; Human Coronaviruses: A Review of Virus–Host Interactions; Published: 25 July 2016

3.      Hector retamal/afp/getty. The city of Wuhan, China, is on lockdown in an attempt to halt the coronavirus outbreak. Rolf hilgenfeld. Volume 557, 30 January2020.

4.      https://www.who.int/news-room/detail/28-01-2020-who-china-leaders-discuss-next-steps-in-battle-against-coronavirus-outbreak Novel Coronavirus(2019-nCoV) Situation Report – 8;

Data as reported by 28 January 2020

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Received on 26.03.2020       Modified on 17.04.2020

Accepted on 11.05.2020      ©AandV Publications All right reserved

Research J. Science and Tech. 2020; 12(2): 147-156.

DOI: 10.5958/2349-2988.2020.00019.4