Rapid differential detection of subtype H1 and H3 swine influenza viruses using a TaqMan-MGB-based duplex one-step real-time RT-PCR assay

Chen, Kaibiao; Kong, Ming; Liu, Jiao; Jiao, Jun; Zeng, Zixiong; Shi, Lijian; Bu, Xinxin; Yan, Yayao; Gao, Ruyi; Liu, Xiaowen; Wang, Xiaoquan; Hu, Jiao; Hu, Shunlin; Jiao, Xinan; Gu, Minghong

doi:10.1007/s00705-021-05127-6

Cited by 4 publications

(7 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In applications where an influence of mismatches was studied, the paired sequence (either a 3ʹ-BHQ1 oligonucleotides or minus strand of dsDNA template in PCR) had an antisense orientation and thus a mismatch CC and AA was present in case of H63D C > G and S65C A > T mutation, respectively. Based on the screening in solution done with the whole series 1 – 12 (see below), three highly active acridines ( 2 , 3 , and 4 ), and one inactive acridine ( 11 ) were selected for modification of the probes. However, the labelling of probes by acridine 3 was not successful even after several repetitions of the reaction.…”

Section: Resultsmentioning

confidence: 99%

“…Preparation of 9-[(6-azidohexyl)amino]-N-(2-guanidinoethyl)acridine-4-carboxamide ( 4 ). Compound 4-Boc (150 mg, 0.23 mmol) was dissolved in DCM (3 ml).…”

Section: Methodsmentioning

confidence: 99%

“…However, the short probes suffer from low melting temperatures and the use of agents thermally stabilising the duplex is mostly unavoidable. Different types of stabilising agents have been described with minor groove binders ( 3 , 4 ), polyamines ( 5 , 6 ) and intercalators ( 7–10 ) being the most promising. Among these agents, acridine derivatives representing the last group, have been demonstrated to be the promising structural motifs thanks to their strong interaction with DNA.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Thermal stabilisation of the short DNA duplexes by acridine-4-carboxamide derivatives

Kostelansky

Miletín

Havlínová

et al. 2022

Nucleic Acids Research

View full text Add to dashboard Cite

The short oligodeoxynucleotide (ODN) probes are suitable for good discrimination of point mutations. However, the probes suffer from low melting temperatures. In this work, the strategy of using acridine-4-carboxamide intercalators to improve thermal stabilisation is investigated. The study of large series of acridines revealed that optimal stabilisation is achieved upon decoration of acridine by secondary carboxamide carrying sterically not demanding basic function bound through a two-carbon linker. Two highly active intercalators were attached to short probes (13 or 18 bases; designed as a part of HFE gene) by click chemistry into positions 7 and/or 13 and proved to increase the melting temperate (Tm) of the duplex by almost 8°C for the best combination. The acridines interact with both single- and double-stranded DNAs with substantially preferred interaction for the latter. The study of interaction suggested higher affinity of the acridines toward the GC- than AT-rich sequences. Good discrimination of two point mutations was shown in practical application with HFE gene (wild type, H63D C > G and S65C A > C mutations). Acridine itself can also serve as a fluorophore and also allows discrimination of the fully matched sequences from those with point mutations in probes labelled only with acridine.

show abstract

Section: Resultsmentioning

confidence: 99%

“…Preparation of 9-[(6-azidohexyl)amino]-N-(2-guanidinoethyl)acridine-4-carboxamide ( 4 ). Compound 4-Boc (150 mg, 0.23 mmol) was dissolved in DCM (3 ml).…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Thermal stabilisation of the short DNA duplexes by acridine-4-carboxamide derivatives

Kostelansky

Miletín

Havlínová

et al. 2022

Nucleic Acids Research

View full text Add to dashboard Cite

show abstract

“…During monthly surveillance of swine IAVs in China during October-December 2020, we collected a total of 376 nasal swab samples from apparently healthy pigs in a slaughterhouse accommodating swine from neighboring regions (Jiangsu, Shandong, and Anhui Provinces in eastern China). We detected H1 subtype swine influenza virus in 9 of those by real-time reverse transcription quantitative PCR; 2 were confirmed as hemagglutinin (HA) positive after inoculating into MDCK cells (12). We further evaluated these 2 swine IAV isolates, A/swine/Jiangsu/HD11/2020 (H1N1) [HD11] and A/swine/Anhui/HD21/2020 (H1N1) [HD21], for their genetic and biologic characteristics.…”

Section: The Studymentioning

confidence: 99%

“…‡Three 6-week-old BALB/c mice per group challenged with 10 6.0 EID50 virus in 50 μL volume were euthanized to collect tissue samples including the lung, turbinate, and brain for virus titration on 3 and 5 d postinoculation. The viral load expressed with virus copies in tissue homogenates was measured through the real-time quantitative reverse transcription PCR method as described (12) embryos with virus titers per 0.1mL >9 log 10 50% egg infectious dose (EID 50 ) (Table 2). However, HD11 replicated much better than HD21 in MDCK cells through the titration of the 50% culture infectious dose (TCID 50 ) value and virus growth at 12-hours intervals across 12-60 hours postinfection (hpi).…”

Section: The Studymentioning

confidence: 99%

Zoonotic Threat of G4 Genotype Eurasian Avian-Like Swine Influenza A(H1N1) Viruses, China, 2020

Gu¹,

Chen²,

Ge³

et al. 2022

Emerg. Infect. Dis.

Self Cite

View full text Add to dashboard Cite

P igs have long been considered a crucial genetic mixing vessel for influenza A viruses (IAVs) of different hosts (1) because of the dual expression of human (SAα-2,6Gal) and avian (SAα-2,3Gal) viral receptors on their respiratory epithelium. Swine IAVs such as H1N1 and H3N2 subtypes sporadically infect humans and are prone to cause bidirectional interspecies transmission at the swine-human interface (2-5). So far, Eurasian avian-like (EA) H1N1 has dominated prevalence in pig herds in China and caused >10 human infections (6-9). In particular, the dominant genotype 4 (G4) EA H1N1 containing 2009 pandemic influenza A(H1N1) polymerase basic (PB) 1 and 2, polymerase acid (PA), nucleoprotein (NP), and matrix (M) genes, plus the triple-reassortant (TR) nonstructural (NS) gene, is thought to be a candidate virus of potential pandemic (10,11). Indeed, a case of human infection with G4 EA H1N1 was reported in Yunan Province, China, in 2021 (8). It is imperative to conduct surveillance on swine IAVs and evaluate their risk to public health.

show abstract

An overview of influenza A virus detection methods: from state-of-the-art of laboratories to point-of-care strategies

Peng,

Wang,

Xie

et al. 2024

Anal. Methods

View full text Add to dashboard Cite

show abstract

Rapid differential detection of subtype H1 and H3 swine influenza viruses using a TaqMan-MGB-based duplex one-step real-time RT-PCR assay

Cited by 4 publications

References 27 publications

Thermal stabilisation of the short DNA duplexes by acridine-4-carboxamide derivatives

Thermal stabilisation of the short DNA duplexes by acridine-4-carboxamide derivatives

Zoonotic Threat of G4 Genotype Eurasian Avian-Like Swine Influenza A(H1N1) Viruses, China, 2020

An overview of influenza A virus detection methods: from state-of-the-art of laboratories to point-of-care strategies

Contact Info

Product

Resources

About