Genetic characterization of HIV-1 viruses among cases with antiretroviral therapy failure in Suzhou City, China.

BACKGROUND: This retrospective study aimed to characterize the distribution of HIV-1 genotypes and the prevalence of drug resistance mutations in people with antiretroviral treatment (ART) failure in Suzhou City, China. METHODS: Pol gene of HIV-1 viruses in blood samples of EDTA anticoagulants from 398 patients with failed antiviral treatment was successfully amplified by using an in-house assay. Drug resistance mutations were analyzed by using the Stanford HIV Drug Resistance Database system ( https://hivdb.stanford.edu/hivdb/by-mutations/ ). HIV-1 genotypes were determined by the REGA HIV subtyping tool (version 3.46, https://www.genomedetective.com/app/typingtool/hiv ). Near full-length genomes (NFLG) of HIV-1 viruses were obtained by next generation sequencing method. RESULTS: Sequences analysis of the pol gene revealed that CRF 01_AE (57.29%, 228/398) was the dominant subtype circulating in Suzhou City, followed by CRF 07_BC (17.34%, 69/398), subtype B (7.54%, 30/398), CRF 08_BC (6.53%, 26/398), CRF 67_01B (3.02%, 12/398) and CRF55_01B (2.51%, 10/398). The overall prevalence of drug-resistant mutations in cases with ART failure was 64.57% (257/398), including 45.48% (181/398) for nucleotide reverse transcriptase inhibitors (NRTIs) mutations, 63.32% (252/398) for non-nucleoside reverse transcriptase inhibitors (NNRTIs) mutations, and 3.02% (12/398) for protease inhibitors (PIs) mutations. Ten near full-length genomes (NFLG) of HIV-1 viruses were identified, including six recombinants of CRF 01_AE and subtype B, two recombinants of CRF 01_AE, subtype B and subtype C sequences, one recombinant of CRF 01_AE and subtype C and one recombinant of CRF 01_AE, subtype A1 and subtype C. CONCLUSIONS: The high prevalence of drug-resistant HIV-1 viruses was a serious challenge for HIV prevention and treatment of people with HIV infection. Treatment regimens for ART failure patients should be adjusted over time based on the outcome of drug resistance tests. NFLG sequencing facilitates the identification of new recombinants of HIV-1.

Retrospective analysis of HIV-1 drug resistance mutations in Suzhou, China from 2009 to 2014.

In this study, we investigated drug resistance levels in human immunodeficiency virus (HIV)-1-infected patients in Suzhou by retrospectively analyzing this property and the characteristics of circulating HIV-1 strains collected from 2009 to 2014. A total of 261 HIV-1-positive plasma samples, confirmed by the Suzhou CDC, were collected and evaluated to detect HIV-1 drug resistance genotypes using an in-house method. The pol gene fragment was amplified, and its nucleic acid sequence was determined by Sanger sequencing. Drug resistance mutations were then analyzed using the Stanford University HIV resistance database ( https://hivdb.stanford.edu ). A total of 216 pol gene fragments were amplified and sequenced with 16.7% (36/216) of sequences revealing these mutations. The drug resistance rates of protease, nucleoside reverse transcriptase, and non-nucleoside reverse transcriptase inhibitors (NNRTIs) were 4/36 (11.1%), 2/36 (5.6%), and 30/36 (83.3%), respectively. Five surveillance drug resistance mutations were found in 36 sequences, of which, three were found among specimens of men who have sex with men. Potential low-level resistance accounted for 33% of amino acid mutations associated with NNRTIs. Two of the mutations, M230L and L100I, which confer a high level of resistance efavirenz (EFV) and nevirapine (NVP) used as NNRTIs for first-line antiretroviral therapy (ART), were detected in this study. Therefore, when HIV-1 patients in Suzhou are administered fist-line ART, much attention should be paid to the status of these mutations that cause resistance to EVP, EFV, and NVP.

Epidemiological and genetic characteristics of the fifth avian influenza A(H7N9) wave in Suzhou, China, from October 2016 to April 2017.

Human infections with H7N9 viruses can cause severe pneumonia and even death. To characterize the epidemiology and genetics of the H7N9 viruses circulating during from October 2016 to April 2017 in Suzhou, China, all pharyngeal swab samples were collected from severe acute respiratory infections (SARI) cases during this fifth wave of infection, and we amplified the H7N9 H7 and N9 genes using a real-time polymerase chain reaction (PCR). Positive samples were subjected to virus isolation and gene sequencing to analyze the evolution and variation of the H7N9 strains. The epidemiological features of H7N9 patients have not changed and there were no significant mutations in the key sites of the hemagglutinin (HA) gene sequence, but we identified the K526R and E627 K substitutions in the PB2 protein. In the neuraminidase (NA) protein, drug-resistant mutations (R294 K and H276Y) occurred in a few strains. Most of the H7N9 viruses isolated from Suzhou had no drug resistance mutations, but it is necessary to closely monitor and analyze the probable emergence of mutations and the spread of resistant strains. The reduction of the N-glycosylation site at position 42 of NA was observed in some strains.

Viral etiology of medically attended influenza-like illnesses in children less than five years old in Suzhou, China, 2011-2014.

Limited information is available on the non-influenza etiology and epidemiology of influenza-like illness (ILI) in China. From April 2011 to March 2014, we collected oropharyngeal swabs from children less than 5 years of age with symptoms of ILI who presented to the outpatient departments of Suzhou University Affiliated Children's Hospital (SCH). We used reverse transcription polymerase chain reaction (rt-PCR) or PCR to detect 11 respiratory viruses. Among 3,662 enrolled ILI patients, 1,292 (35.3%) tested positive for at least one virus. Influenza virus (16.9%) was detected most frequently (influenza A 7.4%, influenza B 9.5%), followed by respiratory syncytial virus (RSV) (5.6%), parainfluenza virus (PIV) types 1-4 (4.8%), human bocavirus (HBoV) (3.8%), human metapneumovirus (HMPV) (3.5%), and adenovirus (ADV) (3.0%). Co-infections were identified in 108 (2.9%) patients. Influenza virus predominantly circulated in January-March and June-July. The 2013-2014 winter peaks of RSV and influenza overlapped. Compared with other virus positive cases, influenza positive cases were more likely to present with febrile seizure, and RSV positive cases were more likely to present with cough and wheezing, and were most frequently diagnosed with pneumonia. These data provide a better understanding of the viral etiology of ILI among children less than 5 years of age in Suzhou, China. Influenza is not only the most frequently identified pathogen but it is also the only vaccine preventable illness among the 11 pathogens tested. Such findings suggest the potential value of exploring value of influenza vaccination among this influenza vaccination target group. J. Med. Virol. 88:1334-1340, 2016. © 2016 Wiley Periodicals, Inc.

Sustained high HIV incidence among men who have sex with men in Jiangsu province, China: based on the limiting-antigen avidity EIA method and six consecutive surveys, 2016-2021.

Background: The epidemic of HIV infection among men who have sex with men (MSM) is a major public health concern in some parts of China, but data on trends in HIV incidence are limited. This study aimed to examine the trends in HIV incidence and factors associated with recent HIV infection among MSM in Jiangsu province, China, based on the limiting-antigen avidity enzyme immunoassay (LAg-Avidity-EIA) method. Methods: Six consecutive surveys were implemented among MSM throughout Jiangsu province from 2016 to 2021. Participants were recruited in three ways. Socio-demographic and behavioral information were collected through face-to-face interviews. Venous blood samples were taken to test for HIV and syphilis. HIV incidence was estimated using the LAg-Avidity-EIA method. Chi-square trend tests were used to observe trends over the years. Multivariate regression analyses were used to identify factors associated with recent HIV infection. Results: A total of 15,401 participants were enrolled in the study. The prevalence of HIV infection ranged from 8.0 to 9.8%, with no consistent rise or fall over the years (P = 0.189). HIV incidence ranged from 5.0 to 9.0%, and no uptrend or downtrend was shown (P = 0.418). MSM who lived locally for more than 2 years (aOR = 1.366, P = 0.019), had a lack of comprehensive HIV knowledge (aOR = 1.643, P = 0.031), had engaged in unprotected anal intercourse (UAI) in the past 6 months (aOR = 7.373, P < 0.001), had been tested for HIV within 12 months (aOR = 1.292, P = 0.035), and tested positive for syphilis (aOR = 2.840, P < 0.001) were likely to be recently infected with HIV. Conclusions: HIV incidence among MSM has remained at a high level in Jiangsu province. In China, health education, condom use, and HIV/syphilis testing should continue to be top priorities for HIV prevention among MSM to reduce HIV transmission.

Real-world data in late presentation of HIV infection in Suzhou, China: Results from four consecutive cross-sectional surveys, 2017-2020.

Objectives: This study aimed to examine the real prevalence of late presentation of HIV infection and to identify factors associated with late HIV presentation among patients with newly diagnosed HIV/AIDS in Suzhou, China. Methods: Patients with newly diagnosed HIV/AIDS who registered in national AIDS surveillance system from 2017 to 2020 were included in this study. Late presentation (LP) of HIV infection was defined as HIV diagnosis with a CD4 count < 350 cells/µL or an AIDS-defining event. Multivariable logistic regression analyses were used to identify factors associated with LP. Results: A total of 2,300 patients were enrolled. 1,325 were classified as late presenters, showing a high percentage of 57.6% (95% CI: 54.5-60.7%) and a rise (P = 0.004) over the four-year period. Patients with newly diagnosed HIV/AIDS who were older than 24 years of age (aOR = 1.549, P = 0.001 for 25-39 years; aOR = 2.389, P < 0.001 for 40 years and older), were Suzhou registered residents (aOR = 1.259, P = 0.026), and were from inpatient and outpatient (aOR = 1.935, P < 0.001) were more likely to be late presentation. Conclusions: This study showed a high percentage and a rise of late presentation of HIV infection among patients with newly diagnosed HIV/AIDS in Suzhou, China, which is a challenge for future prevention and control of AIDS. Targeted measures should be urgently implemented to reduce late HIV diagnosis.

Genetic Characterization of a Novel Reassortant H5N6 Avian Influenza Virus Identified from a 10-Year-Old Girl.

A 10-year-old girl was confirmed, by laboratory tests, to be infected with the H5N6 subtype of the highly pathogenic avian influenza virus in Suzhou city in November, 2018 (JSSZ01). The hemagglutinin (HA) gene of this H5N6 virus belonged to the 2.3.4.4 H5 clade, and the HA linkage peptide of the JSSZ01 strain was RERRRKR↓GLF with multiple basic amino acids. Q226L and G228S mutations were not observed, but S128P, S137A, and T160A substitutions were identified in the receptor binding sites. The resistance mutation D198N in the neuraminidase (NA) protein was also identified in this strain. Additionally, an 11 amino acid (AA positions 59-69) deletion was identified in the NA stalk region. Most genes of JSSZ01 exhibited highest identity with previously characterized H5N6 viruses, but its PA segment sequence was highly similar to previously identified avian H3 viruses (Accession Number: EPI596567 or EPI590058), indicating that JSSZ01 may be created by gene reassortment.

[Genetic characterization of haemagglutinin of novel H1N1 influenza viruses in Suzhou City].

OBJECTIVE: To investigate the genetic variation and molecular characteristics of HA gene of influenza A (H1N1) virus isolated in Suzhou during from2009 to 2011. METHODS: Viral RNA of 5 Suzhou isolates was isolated and their HA gene were amplified and sequenced by the primers and protocol recommended by WHO, and the sequences together with other sequences downloaded from GenBank were analyzed by several bioinformatics software. RESULTS: Compared with vaccine strain, the average homogeneity of nucleotide and amino acids of 5 Suzhou isolates were between 98.8-99.4% and 98.8-99.4% respectively. All of the 5 strains have 1 amino acids replacement in Sb region and 2 strains have 2 amino acids replacement in Ca region. Strains from and outside Suzhou both showed a trend of clustering by collection year. CONCLUSION: The Suzhou strains are still in stable condition although 1-2 amino acids replacement had happened in antigenic sites.

[The application of molecular differential diagnoses platform in identification of the respiratory pathogens].

AIM: To establish a MDD (molecular differential diagnoses) platform for diagnosing the pathogens that may cause respiratory infection by combination of the advanced Tem-PCR(Target enriched multiplex PCR)with xMAP(multiple analyses profiling), and to evaluate the reliability and further use the platform to test clinic samples. METHODS: 22 throat swab specimen from outpatient patients of respiratory department in First Affiliated Hospital of Suzhou University and 20 respiratory tract lavage fluid specimen from inpatients of respiratory department in Affiliated Children Hospital of Suzhou University were collected, and the nucleic acids of the samples were amplified by Tem-PCR and xMAP. RESULTS: Testing of the the known samples showed that the platform had excellent specificity and sensitivity. Testing of the clinic samples showed that the positive rate of the respiratory tract lavage fluid specimen was 63.6%, higher than that of the throat swab specimen, and that the positive rate of RNA pathogens was higher than that of DNA pathogens. CONCLUSION: A reliable MDD platform for detection of respiratory pathogens has been established with high-throughput detection capacity.