Detection of D151G/N mutations in the neuraminidase gene of influenza A (H3N2) viruses by real‐time RT‐PCR allelic discrimination assay

Zadeh, Vahid Rajabali; Jagadesh, Anitha; Krishnan, Anjana; Arunkumar, Govindakarnavar

doi:10.1002/jmv.24757

Cited by 3 publications

(2 citation statements)

References 10 publications

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“…For the different results of clinical specimens by CMT and FMT, an influenza A real-time RT-PCR detection kit was also used to detect influenza A again [21]. The results revealed that 26 of the 33 specimens that detected negative with CMT but positive with FMT, detected positive with the real-time RT-PCR detection method, and none of the 3 specimens that had previously detected positive with CMT but negative with FMT provided a positive result (Table 5).…”

Section: Resultsmentioning

confidence: 99%

Comparison of the detection performance of two different one-step-combined test strips with fluorescent microspheres or colored microspheres as tracers for influenza A and B viruses

Liao

Wang

et al. 2019

Virol J

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Background Influenza A and B viruses mainly cause respiratory infectious disease. Till now, few tests are able to simultaneously detect both, especially in primary medical establishments. Methods This study was designed to compare the performance of two different one-step-combined test strips for the detection of influenza A and B: one strip with fluorescent microspheres for tracers (FMT); and the other strip with colored microspheres for tracers (CMT). To test the strips, cultures of influenza A, B, and other pathogenic viruses were used, in addition to 1085 clinical specimens from symptomatic patients with respiratory infections. Real-time RT-PCR was also considered as a reference method used to detect the different results of FMT and CTM. Results Detection thresholds for influenza A and B cultures using serial dilutions revealed that the sensitivity of FMT was higher than that of CMT (both P < 0.05). With the culture mixtures of Coxsackie virus (A16), enteric cytopathic human orphan virus (ECHO type30), enterovirus (EV71), rotavirus (LLR strain), and enteric adenovirus (AdV 41), specificity assessment demonstrated that there was no cross reaction during the usage of the two test strips as shown by the results which were negative. In the detection of influenza A in 1085 clinical specimens, the total coincidence rate was 96.7%, the positive coincidence rate was 97.1%, and the negative coincidence rate was 96.7%. In the case of influenza B detection, the total coincidence rate was 99.1%, the positive coincidence rate was 92.6%, and the negative coincidence rate was 98.5%. In addition, with influenza A or B real-time RT-PCR detection method, the results showed that, for influenza A, 26 of the 33 specimens that negative with CMT but positive with FMT, showed positive results, and none of the 3 specimens that positive with CMT but negative with FMT showed a positive result; For influenza B, 12 of the 15 specimens that negative with CMT but positive with FMT, showed positive results, and none of the 5 specimens that positive with CMT but negative with FMT showed a positive result. Conclusions FMT performed better than CMT in the combined detection of influenza A and B viruses.

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Section: Resultsmentioning

confidence: 99%

Comparison of the detection performance of two different one-step-combined test strips with fluorescent microspheres or colored microspheres as tracers for influenza A and B viruses

Liao

Wang

et al. 2019

Virol J

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“…For instance, HA Q226L [19][20][21] and G186V [19,[22][23][24] mutations were shown to increase the affinitive binding between H7N9 viruses and human-type receptors. Moreover, recent studies showed that N2 with a D151G amino acid substitution [25][26][27], or N1 with a G147R amino acid substitution [26,28,29], had similar properties of enhanced affinitive binding for SA receptors. However, further investigation is required to explore whether the pathology of H7N9 infections could be caused by other amino acid substitutions, resulting from unknown conformational variations in dual-receptor binding capacity of the H7 protein, or additional affinity owing to the N9 protein.…”

Section: Introductionmentioning

confidence: 99%

Identifying novel amino acid substitutions of hemagglutinin involved in virulence enhancement in H7N9 virus strains

Zhang

Guo

et al. 2021

Virol J

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Background To identify site-specific features of amino acid substitutions that confer enhanced H7N9 virulence in humans, we independently generated mammalian-adapted variants of A/Anhui/1/2013 (AH-H7N9) and A/Shanghai/2/2013 (SH-H7N9) by serial passaging in Madin-Darby canine kidney (MDCK) cells. Methods Virus was respectively extracted from cell culture supernatant and cells, and was absolutely quantified by using real-time polymerase chain reaction. Viral RNAs were extracted and subjected to sequencing for identifying mutations. Then, site-specific mutations introduced by viral passaging were selected for further constructing HA7 or NA9 mutant plasmids, which were used to generate recombinant viruses. The interaction between the recombinant HA and receptors, H7N9-pseudotyped viruses and receptors were detected. Results Both subtypes displayed high variability in replicative capability and virulence during serial passaging. Analysis of viral genomes revealed multiple amino acid mutations in the hemagglutinin 7 (HA7) (A135T [AH-H7N9], T71I [SH-H7N9], T157I [SH-H7N9], T71I-V223I [SH-H7N9], T71I-T157I-V223I [SH-H7N9], and T71I-T157I-V223I-T40I [SH-H7N9]), and NA9 (N171S [AH-H7N9] and G335S [AH-H7N9]) proteins in various strains of the corresponding subtypes. Notably, quite a few amino acid substitutions indeed collectively strengthened the interactions between H7N9 strains and sialic acid receptors. Moreover, some of the amino acid substitutions identified were highly and specifically cytopathogenic to MDCK cells. Conclusions This study demonstrated that AH-H7N9 and SH-H7N9 subtypes can acquire enhanced receptor affinity for sialic receptors through novel amino acid substitutions. Such changes in affinitive interactions are conferred by site-specific mutations of HA7 proteins that affect the virulence and pathology of the virus strain, and/or limited compatibility between the host and the virus strain.

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Key amino acid residues of neuraminidase involved in influenza A virus entry

Chen

et al. 2019

Pathogens and Disease

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Generally, influenza virus neuraminidase (NA) plays a critical role in the release stage of influenza virus. Recently, it has been found that NA may promote influenza virus to access the target cells. However, the mechanism remain unclear. Here, we reported that peramivir indeed possessed anti-influenza A virus (IAV) activity in the stage of viral entry. Importantly, we verified the critical residues of influenza NA involved in the viral entry. As a result, peramivir as an efficient NA inhibitor could suppress the initiation of IAV infection. Furthermore, mutational analysis showed NA might be associated with viral entry via amino acids residues R118, E119, D151, R152, W178, I222, E227, E276, R292 and R371. Our results demonstrated NA must contain the key amino acid residues can involve in IAV entry.

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Detection of D151G/N mutations in the neuraminidase gene of influenza A (H3N2) viruses by real‐time RT‐PCR allelic discrimination assay

Cited by 3 publications

References 10 publications

Comparison of the detection performance of two different one-step-combined test strips with fluorescent microspheres or colored microspheres as tracers for influenza A and B viruses

Comparison of the detection performance of two different one-step-combined test strips with fluorescent microspheres or colored microspheres as tracers for influenza A and B viruses

Identifying novel amino acid substitutions of hemagglutinin involved in virulence enhancement in H7N9 virus strains

Key amino acid residues of neuraminidase involved in influenza A virus entry

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