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Influenza viruses resistant to oseltamivir, news and updates
#148, Kent.N. "ProMED: ... Laboratory tests on 6 samples of the H5N1 strain of avian influenza collected from poultry in Indonesia in 2005 showed they were between 15 and 30 times less sensitive to Tamifluthan a family of the viruses collected in Southeast Asia a year earlier."
Does the above means results from poultry lab. experiments provenience,
or inc. folks treated infected farming poultry with Tamiflu antivirals and those poultry exibited than the above lab results of resistance?
The antiviral resiatnce was reported at the Options VI meeting over a year ago (which means they had the data almost 2 years ago). These results would be for testing of H5N1 from samples. There was no specific change (like H274Y) in the isolates.
The only human H5N1 sequence I know of is from the Karo patient who stopped taking his Tamiflu. He died and H274Y was in his H5N1 sequence.
Tamiflu is dispersed like candy in Indonesia, and the last public human sequence was from January, 2007.
It is likely that there is unreported H274Y in H5N1 and H1N1 in Indonesia.
Dr Niman,
Thank you for spending time on my questions. Obviously I have a lot to learn with regards to sequencing and interpreting NCBI data. The data you are relying on seems to be contributed by two groups. One that does not mention zanamivir in their submissions (as I noted in an above post) and the other group who seem to be submitting a mix of influenza data with no mention of zanamivir and a title for an as yet unpublished paper. The sooner they are published, the sooner we'll (I'll) know degrees of resistance, viability of the resistant strain etc.
I would have thought this would have been a priority for the medical community, an opportunity to say Relenza is as big a dud as Tamiflu (as happened with the behaviour warning from Japan).
LOCUS CY030873 1396 bp cRNA linear VRL 17-MAR-2008
DEFINITION Influenza A virus (A/Thailand/39/2008(H1N1)) segment 6 sequence.
ACCESSION CY030873
VERSION CY030873.1 GI:170026929
KEYWORDS .
SOURCE Influenza A virus (A/Thailand/39/2008(H1N1))
ORGANISM Influenza A virus (A/Thailand/39/2008(H1N1))
Viruses; ssRNA negative-strand viruses; Orthomyxoviridae;
Influenzavirus A.
REFERENCE 1 (bases 1 to 1396)
AUTHORS Hurt,A.C., Kelso,A. and Barr,I.G.
TITLE Community cases of zanamivir-resistant human influenza viruses with
a novel mutation in the neuraminidase gene
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 1396)
AUTHORS Hurt,A.C., Deng,Y.-M. and Komadina,N.
TITLE Direct Submission
JOURNAL Submitted (14-MAR-2008) WHO Collaborating Centre for Reference and
Research on Influenza, 45 Poplar Rd, Parkville, Victoria 3052,
Australia
FEATURES Location/Qualifiers
source 1..1396
/organism="Influenza A virus (A/Thailand/39/2008(H1N1))"
/mol_type="viral cRNA"
/strain="A/Thailand/39/2008(H1N1)"
/serotype="H1N1"
/isolation_source="gender:M; age:23"
/specific_host="Human"
/db_xref="taxon:511421"
/segment="6"
/lab_host="MDCKX passage(s)"
/country="Thailand"
/collection_date="02-Jan-2008" gene 1..>1396
/gene="NA" CDS 1..>1396
/gene="NA"
/codon_start=1
/product="neuraminidase"
/protein_id="ACB05991.1"
/db_xref="GI:170026930"
/translation="MNPNQKIITIGSISIAIGIISLMLQIGNIISIWASHS IQTGSQN
NTGICNQRIITYENSTWVNHTYVNINNTNVVAGEDKTSVTLAGNSSLCSI SGWAIYTK
DNSIRIGSKGDVFVIREPFISCSHLECRTFFLTKGALLNDKHSNGTVKDR SPYRALMS
CPLGEAPSPYNSKFESVAWSASACHDGMGWLTIGISGPDNGAVAVLKYNG IITGTIKS
WKKQILRTQESECVCMNGSCFTIMTDGPSNKAASYKIFKIEKGKVTKSIE LNAPNFHY
EECSCYPDTGIVMCVCRDNWHGSNRPWVSFNQNLDYQIGYICSGVFGDNP RPEDGEGS
CNPVTVDGANGVKGFSYKYDNGVWIGRTKSNRLRKGFEMIWDPNGWTNTD SDFSVKQD
VVAITDWSGYSGSFVQHPELTGLDCIRPCFWVELVRGLPRENTTIWTSGS SISFCGVN
SDTANWSWPDGAELP"
ORIGIN
1 atgaatccaa atcaaaaaat aataaccatt ggatcaatca gtatagcaat cggaataatt
61 agtctaatgt tgcaaatagg aaatattatt tcaatatggg ctagtcactc aatccaaact
121 ggaagtcaaa acaacactgg aatatgcaac caaagaatca tcacatatga aaacagcacc
181 tgggtgaatc acacatatgt taatattaac aacactaatg ttgttgcagg agaggacaaa
241 acttcagtga cattggccgg caattcgtct ctttgttcta tcagtggatg ggctatatac
301 acaaaagaca acagcataag aattggctcc aaaggagatg tttttgtcat aagagaacct
361 ttcatatcat gttctcactt ggaatgcaga accttttttc tgaccaaagg cgctctatta
421 aatgacaaac attcaaatgg gaccgtaaag gacagaagtc cttatagggc cttaatgagc
481 tgtcctctag gtgaagctcc gtccccatac aattcaaagt tcgaatcagt tgcatggtca
541 gcaagcgcat gccatgatgg catgggctgg ttaacaatcg gaatttctgg tccagacaat
601 ggagctgtgg ctgtactaaa atacaacgga ataataactg gaaccataaa aagttggaaa
661 aagcaaatat taagaacaca agagtctgaa tgtgtctgta tgaacgggtc atgtttcacc
721 ataatgaccg atggcccgag taataaggcc gcctcgtaca aaattttcaa gatcgaaaag
781 gggaaggtta ctaaatcaat agagttgaat gcacccaatt ttcattatga ggaatgttcc
841 tgttacccag acactggcat agtgatgtgt gtatgcaggg acaactggca tggttcaaat
901 cgaccttggg tgtcttttaa tcaaaacttg gattatcaaa taggatacat ctgcagtgga
961 gtgttcggtg acaatccgcg tcccgaagat ggagagggca gctgcaatcc agtgactgtt
1021 gatggagcaa acggagtaaa agggttttca tacaaatatg ataatggtgt ttggatagga
1081 aggaccaaaa gtaacagact tagaaagggg tttgagatga tctgggatcc taatggatgg
1141 acaaataccg acagtgattt ctcagtgaaa caggatgttg tagcaataac tgattggtca
1201 gggtacagcg gaagtttcgt ccaacatcct gagttaacag gattggactg tataagacct
1261 tgcttctggg ttgagttagt cagagggctg cctagagaaa atacaacaat ttggactagt
1321 gggagcagca tttctttttg tggcgttaat agtgatactg caaactggtc ttggccagac
1381 ggtgctgagt tgccat
Dr Niman,
Thank you for spending time on my questions. Obviously I have a lot to learn with regards to sequencing and interpreting NCBI data. The data you are relying on seems to be contributed by two groups. One that does not mention zanamivir in their submissions (as I noted in an above post) and the other group who seem to be submitting a mix of influenza data with no mention of zanamivir and a title for an as yet unpublished paper. The sooner they are published, the sooner we'll (I'll) know degrees of resistance, viability of the resistant strain etc.
I would have thought this would have been a priority for the medical community, an opportunity to say Relenza is as big a dud as Tamiflu (as happened with the behaviour warning from Japan).
Abstract of CDC presentation at Options VI
Abstract O66
Monitoring of Influenza Virus
Susceptibility to Neuraminidase
Inhibitors (NAIs)
LV Gubareva, VM Deyde, X Xu, RA Bright
*, TG Sheu, AI Klimov
Influenza Division, National Center for Immunization and Respiratory Diseases,
Centers for Disease Control and Prevention, Atlanta Georgia, USA
*Present address:
Novavax, One Taft Court, Suite 200 Rockville, Maryland 20850 USA
Oseltamivir and zanamivir are currently approved NAIs for
the control of influenza infections; a third drug, peramivir, is
undergoing clinical evaluation. NAI-susceptibility is commonly
assessed through an enzyme activity inhibition assay with either
chemiluminogenic or fluorogenic substrate. Standardization
of assay conditions is essential for reliable drug resistance
monitoring. As a part of continued influenza strain surveillance,
we utilized a recently developed NAStar kit for the detection
of NAI-resistant mutants by means of chemiluminescence.
Zanamivir and oseltamivir were used to determine IC
50 values
for 133 A(H1N1), 186 A(H3N2), and 118 B viruses collected in
2005-2007. All viruses appeared sensitive to both drugs with
one exception. An influenza B virus, with a R371K substitution
at the enzyme active site, exhibited elevated IC50 values when
tested with both zanamivir (~150nM) and oseltamivir (~1,000
nM), which indicates drug resistance. Another influenza B
virus, although sensitive to both drugs, contained substitution
H274Y in the NA active site, which was previously shown to
confer in-vitro resistance to peramivir. The two newly identified
variants and a panel of well-characterized oseltamivir- or
zanamivir-selected mutants and their respective wild types
(WT) were then tested with four NAIs (oseltamivir, zanamivir,
peramivir, and A-315675). When tested with oseltamivir, the
oseltamivir-selected mutants [e.g., H274Y(N1)] exhibited 10-
fold or greater IC50 values than their WT. In contrast, a majority
of the zanamivir-resistant viruses [e.g., E119G(N2)] had <10-
fold increase in IC50 values compared to their WT, when tested
with zanamivir. Nevertheless, when tested with oseltamivir,
two of those mutants [R152K(B) and R292K(N2)] exhibited >10-
fold increase in their IC50 values compared to their WT viruses.
The H274Y(B) variant exhibited a 4-fold increase in IC50 value
for peramivir compared to that of the control virus. When the
same variant was tested in fluorescent assay, increase in IC50
values was greater for both peramivir (~10-fold) and oseltamivir
(~5-fold). The R371K variant exhibited cross-resistance to all
four NAIs in both assays (≥10 fold). A majority of NAI-resistant
mutants were detected with NAStar kit; thus, it could be
useful for drug resistance surveillance. To ensure dependable
detection of every zanamivir- and peramivir-resistant mutant,
Abstract O67
Novel Mutations in the ?150-Cavity?
of N1 Neuraminidase Confer Reduced
Sensitivity to the Neuraminidase Inhibitors
Aeron C Hurt
1,2, Ian G Barr1,2
1
World Health Organisation Collaborating Centre for Reference and Research on
Influenza, Parkville, Victoria 3052, Australia; 2Monash University, School of Applied
Sciences, Churchill, Victoria 3842, Australia
The neuraminidase (NA) inhibitors are a specifically designed
class of antiviral drugs that bind to the active site of the NA
surface glycoprotein of newly formed influenza virus particles
and prevent their efficient release from the host cell. Although
resistance to these inhibitors is relatively rare, investigation
of circulating strains for changes in susceptibility remains
important. Analysis of recent influenza isolates received at
the WHO Collaborating Centre for Influenza in Melbourne has
revealed four A(H1N1) strains with increased IC
50 (50% inhibitory
concentration) values to either zanamivir or oseltamivir
carboxylate. These isolates were received from locations
where little or no NA inhibitors were in use and are therefore
assumed to be derived from patients not being treated with
the NA inhibitors. Two of the strains, which demonstrated a
250-fold increase in zanamivir IC50 but no change in oseltamivir
carboxylate IC50, were found to have a Q136K mutation in the
NA gene, while a third strain which had a 30-fold increase in
zanamivir IC50 and no change in oseltamivir carboxylate IC50
was found to have a K150T mutation in the NA gene. The fourth
A(H1N1) virus identified to confer reduced sensitivity to the NA
inhibitors, had a 30-fold increase in zanamivir IC
50 and a 10-fold
change in oseltamivir carboxylate IC50 and was found to have a
K143R NA gene mutation. Interestingly, all of the four mutants
contained substitutions at residues located in and around
the recently identified ?150-cavity? (Russell et al, 2006), which
following x-ray crystallography was shown to be adjacent to
the NA active site of group-1 neuraminidases (N1, N4, N5, N8),
but is not present in group-2 neuraminidases (N2,N3,N6,N7,N9).
None of these sites has previously been identified as conferring
NA inhibitor resistance either in vitro or in clinical studies. Given
that residues in this cavity appear to impact on zanamivir
sensitivity, and to a lesser extent oseltamivir sensitivity, it will
therefore be important to determine the extent to which both
H1N1 and highly pathogenic H5N1 viruses with mutations in
Abstract O69
Indonesian H5N1 Isolates Demonstrate
Decreased Sensitivity to Oseltamivir
JL McKimm-Breschkin
1, P Selleck2, T Usman3, M Johnson2
1
CSIRO Molecular and Health Technologies, Parkville, Australia; 2CSIRO Livestock
Industries, Geelong, Australia; 3Disease Investigation Centre, Yogjakarta, Indonesia
Two different strains of highly pathogenic H5N1 avian influenza
have been circulating since 2003. Clade 1 has been found in
Vietnam, Thailand, Cambodia, Laos and Malaysia. Clade 2
subsequently emerged and spread from China to Indonesia,
Europe and Africa in 2004-2005. Due to its systemic availability
oseltamivir is the drug of choice for treating infected humans.
We tested the drug sensitivity of NAs from eight H5N1 viruses
from 2004 from Vietnam, two from 2004 from Malaysia, six
from Cambodia from 2004, four from 2005 and six clade 2 2005
viruses from Indonesia. Viruses were isolated from chickens,
ducks, geese and quail. In the absence of a validated cell culture
assay the IC
50 measured in the MUNANA based NA enzyme
inhibition assay was used for measuring drug sensitivity. All
clade 1 and clade 2 viruses had a similar sensitivity to zanamivir
as the reference human H1N1 NA. Sensitivities of the NAs to
oseltamivir fell into three groups compared to the human H1N1
strain. The clade 1 isolates from 2004 were all more sensitive
to oseltamivir than the human H1N1 control. However the NAs
of the 2005 Cambodian viruses showed a 6-7 fold decrease
specifically in oseltamivir sensitivity compared to the 2004
Cambodian isolates. These 2005 isolates came from the same
area as one of the more sensitive 2004 isolates, suggesting
regional evolution had occurred. Of more concern was the
third group. The NAs from all the clade 2 2005 Indonesian
viruses demonstrated a 25-30-fold decrease in sensitivity
specifically to oseltamivir compared to the clade 1 viruses. This
decrease in sensitivity is even greater than that conferred by
the N294S recently detected in Egypt, known to be selected
for by oseltamivir treatment. There was no altered sensitivity
to a third inhibitor, 4-AminoNeu5Ac2en, which shares the
4-amino substitution with oseltamivir, but has the original
glycerol side chain at the 6? position. This indicates that the
altered binding of oseltamivir was due to altered interaction
with its hydrophobic pentyl ether group, thus explaining why
11 around 2007, neuraminidase (NA) 275 proteins from the second histidine tylosin amino acids (H275Y <SUP>*)</SUP> to replace the strong resistance to oseltamivir for influenza A/H1N1 virus subtype, and 67 percent in Norway Led, EU countries, more than 20 percent of the overall high-frequency has to be found in 1).
For this reason, WHO GUROBARUINFURUENZASABEIRANSUNETTOWAKU the global NA inhibitor (NAI) to strengthen surveillance-resistant strain, resistant strain emerged in the country to report the situation said.
This was the week or month from around the world have information in their units, their results of the WHO website on a regular basis (HP) came to be published in the 2).
April-October 2008 of the current frequency in the entire world is 39 percent, from late 2007 until March of this year's 16 percent larger than the Southern Hemisphere countries in the world, including the resistant strain is starting to spread .
In particular, Senegal, South Africa A/H1N1 isolate the resistance of 100 percent and 88 percent in Africa but the entire region is a two-resistant strain).
These shares oseltamivir resistance is not taking patients have been isolated from the normal trend of the market share with virulence as the spread between people, and 3), the flu is a big problem.
Meanwhile, the country of the world's production of oseltamivir for more than 70 percent have clinical use in the field, because in addition to the market trend-resistant strain of the selective pressure of drug-resistant strain of the emergence of high-frequency危惧SA of the world Shares in the incidence of resistance in the country are watching developments.
Against this background, the National Institute of Infectious Diseases (lab infection), the local Public Health Research Institute (research areas) with the cooperation of the 2007/08 season, A/H1N1 strain isolated in the country for emergency surveillance-resistant strain To be implemented.
The progress report in Japan that an emergency surveillance, as the first report on the issue IASR 6 is 4), this time, was later added to isolate the analysis results of the 2007/08 season, including the separation of the total shares As reported in the second report together.
(*)'s H274Y in the various papers have representation, which, H3N2 subtype of the virus NA proteins, amino acids were based on the number notation (N2 notation) and, H1N1 cases in the NA protein , A marker of resistance from the amino acid methionine number at 275 and counting.
よって、本文では耐性マーカーのアミノ酸番号をH275Yで統一する。
Thus, the amino acids in the body and a number of resistance markers in a unified H275Y.
1. Occur the domestic situation
-resistant strain of the emergency-resistant strain of surveillance research obtained in the cooperation received from the State-resistant strain detected by the situation in Table 1 (pdf) to the (virus-resistant The detection method IASR 6 issue the first report of the methods section) 4).
Analysis of 1734 total number of shares of stock during the 45-resistant strain is identified, the frequency of domestic-resistant strain was 2.6 percent (Table 1).
Past look at the frequency incidence, and in 2007 the 331 shares per share (0.3 percent), while 44 stocks in 2008, 1403 shares (3.1 percent), since 2008 the incidence of high frequency.
-Resistant strain was isolated in the 10 prefectures of Honshu (Figure 1 pdf), Tottori Prefecture, which excludes often occur in nine prefectures, respectively, from 1.2 to 7.3 percent.
Meanwhile, shares in Tottori Prefecture during the 22 stocks in the 68-resistant strain, the frequency has been outstanding and 32.4 percent (Figure 1).
These state-resistant strain occurred in the city and another (Figure 2 pdf), with a focus on the city of Tottori Prefecture in late January from the eastern most isolated stocks, NA Hawaii strain on the genetic family tree (the term analysis of the evolutionary tree ) Belongs to, but separate shares in February and March into the Nordic some strains, the two strains were mixed.
On the other hand, with a focus on Kurayoshi from the central prefecture, February and March are resistant to the separation of stocks, they belong to the Nordic system.
From the eastern part of Tottori Prefecture, the prefectural government, in the central prefecture of two different genetic strains resistant strain of the epidemic have been widely suggested.
In Tottori Prefecture, the so-resistant strain was prevalent, despite the high frequency of the neighboring prefecture of the resistant strain of the infected people in the province are low due to age, moving beyond the County One of the few considered to be the cause.
Prevalent in the 2007/08 season, influenza A/H1N1 virus, NA SABUKUREDO on the evolutionary tree of a gene 2B (amino acid marker: H45N, G249K, T287I, K329E, G354D) and SABUKUREDO 2C (amino acid marker: S82P, M188I, I267M, L367I, V393I, T453I) into the Figure 3: upper left corner (pdf)].
2B SABUKUREDO two more within goodbye to the strains, D354G marker in the world and are widely separated Scandinavian-resistant strains of the shares, the markers do not have a vaccine strain A / Brisbane / 59/2007, including strains of Hawaii Resistance was divided into shares.
ハワイ系統の耐性株は分離株数から見ると世界的には少数派である。
Shares of Hawaiian-resistant strains isolated from the number of shares and the world is in the minority.
-Resistant strain isolated in Japan, Hawaii, a lot of strain was recognized as belonging to the city of Yokohama, Tottori and Okayama prefectures from belonging to resistant strains of the Nordic stock is also detected.
15 shares for shares in the resistance, and its share of old and new vaccine for ferrets to see a similar line with anti-serum hemagglutination inhibition (HI) test was conducted to analyze the results of antigen in the country and Nordic strains isolated strains either Hawaii -Resistant strain, the current term of the vaccine strain A / Brisbane / 59/2007 shares were similar (IASR 6 issue, the first report) 4).
このことから、これら耐性株に対して、今冬のワクチンは有効であることが示唆された。
For this reason, these stocks resistant to the vaccine this winter suggested that to be effective.
The separation of the 2007/08 season, 240 shares drug-sensitive stocks (Table 1: blue) drug susceptibility testing for the NAI as a result of the drug oseltamivir-sensitive shares, 50% NA activity inhibition concentration (IC50) in the average value of the 0.09nM .
On the other hand, domestic resistance to oseltamivir IC50 value of the shares, or more generally indicates 30.0nM, drug-sensitive shares compared to 300 times more sensitive to oseltamivir also has been slow.
These Zanamivir-resistant strain to the most sensitive of all, but isolated in Tottori Prefecture per share (A / Tottori / 44/2008), oseltamivir, was resistant to both Zanamivir.
また、別の1株(A/鳥取/16/2008)は、ザナミビルに耐性であった( 表2 )。
Another per share (A / Tottori / 16/2008), was resistant to Zanamivir (Table 2).
On the other hand, belong to the clade Yokohama 2C isolate (A / Yokohama / 91/2007), NAI is a different mechanism of action of anti-flu drug amantadine resistance to the virus marker (M2 protein S31N) and Life Oseltamivir and amantadine-resistant and 2.
5. A/H3N2 and B-NAI-resistant strain of influenza virus infection surveillance
surveillance research for the NAI-resistant strain, A/H3N2 and B-type influenza viruses, the total length of the NA gene analysis and drug susceptibility testing parallel NAI Are carried out.
Table 2, A/H1N1 in addition to the shares in the separation of the 2007/08 season, A/H3N2 and B-type strain was resistant strain found in the summarize the situation.
For the 2007/08 season, A/H1N1 isolate emergency surveillance of the drug-resistant strain, resistant strain of the outbreak in Japan is often lower than in foreign countries, EU countries so far, African countries, like Australia, serious Is not ready.
In addition, A/H3N2 and B-resistant strain of the flu virus had not been confirmed so far in the domestic policies oseltamivir in the treatment of influenza will not be a big influence.
In addition, the separation of the domestic season-resistant strain of the vaccine strain A / Brisbane / 59/2007 on the genetic and antigenic are similar, the vaccine is believed to be valid.
In addition, the two share some exceptions, almost all of the Zanamivir-resistant strain that is sensitive, and effective treatment with Zanamivir.
これまでも、NA阻害剤耐性株はA/H1N1、A/H3N2およびB型インフルエンザウイルスで低頻度(0.11~0.68%)ながら見つかってきたが(IA SR 6月号、第1報、表1 )4)、これらは市中流行株としてヒトからヒトへ伝播拡大することはなく、自然消滅していた。
So far, NA stock inhibitor-resistant A/H1N1, A/H3N2 and B-type influenza virus in the low-frequency (0.11 to 0.68%) have been found and (IASR 6 issue, the First Report, Table 1) 4 ), Which shares the market as a fashion spread from human to human, not to expand natural been extinguished.
However, other countries of the oseltamivir-resistant strain has been found in the continued transmission capacity and market share trends, and in some countries is becoming a major epidemic.
Japan also is not clear why the projection in Tottori Prefecture, and high frequency been observed, this winter to expand the nation feared an epidemic.
Shares of oseltamivir resistance than normal-sensitive stocks and can not accept the differences in clinical symptoms, but then Oseltamivir is a lot for our country, shares of resistance to amantadine-resistant strain of If the epidemic is in the mainstream of shares, Treatment with a drug policy will be a big impact.
In the clinical field, and rapid diagnostic kit of the type A or B type is possible to identify, A/H1 and A/H3 subtype is impossible to identify the emission shares are resistant to the patient To prescribe oseltamivir could.
Thus, even in winter fashions, A/H1N1 and B-shares as well as influenza A/H3N2 virus at the national level, including the need for continued monitoring.
The Third National Institute of Infectious Diseases Department viral drug-resistant strain of influenza surveillance room team Biotechnology Institute of Technology and Evaluation Division headquarters influenza virus genome analysis of the local National Institutes of Health genetic analysis team
Comment