Development of a novel target-enriched multiplex PCR (Tem-PCR) assay for simultaneous detection of five foodborne pathogens

Influenza is a contagious respiratory disease and risks public health in China, and it has caused wide public concern in recent years. Immunocompromised patients, such as children and elderly people, suffer more severe influenza complication and some extreme cases are even life threatening. To identify the influenza characteristics and its correlation with various climatic and environmental pollution factors, we collected the reported influenza epidemic of hospitalized children in Children's Hospital of Soochow University from 2016 to 2019. Our results show that the main influenza virus subtypes are A/H1N1, A/H3N2, B/BV, and B/BY. We also identified the characteristics of the prevalent influenza virus subtypes in different months, seasons, years, and patients' age. Of all the influenza infected patients, the most susceptible groups are children over 3 to 5 years of age, and more cases are reported in winter than other seasons. We also found that influenza is also highly correlated with climatic and environmental pollution factors, and the autoregressive integrated moving average model is employed for the short-term influenza prediction in Suzhou city, which can provide scientific basis for the prevention and control of influenza and public health decision-making.

Epidemiology of influenza in hospitalized children with respiratory tract infection in Suzhou area from 2016 to 2019.

A novel target-enriched multiplex polymerase chain reaction (Tem-PCR) assay for the simultaneous detection of Salmonella spp., Listeria monocytogenes and Escherichia coli O157:H7 was developed in the present study. The invA, hly and rfbE genes were selected as target genes for identifying Salmonella spp., L. monocytogenes and E. coli O157:H7, respectively. Three pairs of target-enriched long primers with a composition of pathogen-specific PCR primer combined with a universal oligonucleotide sequence at 5′ segment of each pathogen-specific PCR primer were designed. Following optimization of reaction conditions, the Tem-PCR assay for detecting Salmonella spp., L. monocytogenes and E. coli O157:H7 was successfully developed, which was able to simultaneously detect the three foodborne pathogens from pure cultures with an analytical detection limit of ca. <110 cfu/mL for each target. The Tem-PCR assay showed a high specificity to the target bacteria without specific priming observed in nontarget bacteria and also showed a potential diagnostic capability in practice. Compared with conventional multiplex PCR assay, the Tem-PCR assay does need not to optimize the concentration of each pair of primers, suggesting an efficient detection method for foodborne pathogens in laboratory diagnosis.

Practical Applications
Applying the target-enriched multiplex polymerase chain reaction assay to 355 food samples, including poultry meat, raw pork, raw milk, egg, sausage, raw beef, milk powder and frozen meat, revealed that 21 food samples were positive in this assay, in accordance with the testing results of conventional culture-based method, indicating a rapid and reliable tool for efficient screening of Salmonella spp., Listeria monocytogenes and Escherichia coli O157:H7 from food.

Target-Enriched Multiplex PCR (Tem-PCR) Assay for Simultaneous Detection of Salmonella spp., Listeria Monocytogenes and Escherichia Coli O157:H7 in Food

Introduction: This study aimed to identify factors associated with late diagnosis and clinically monitor newly diagnosed HIV/AIDS patients. Method: Retrospective study, based on secondary data from a specialized unit at the Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto of the University of Sao Paulo. Data collection included sociodemographic, behavioral, clinical, and laboratory data of newly diagnosed HIV patients between 2015 and 2019. Data analysis was undertaken using inferential statistical tests. Results: A total of 314 individuals were newly diagnosed with HIV/AIDS, 86.6% (272) had a late diagnosis and 53.8% (169) were diagnosed very late. Using the adjusted odds ratio, we observed that bisexual and MSM patients were less likely to have a late diagnosis compared to straight patients. Individuals who entered through the emergency department and Outpatient Clinic had a lower chance of having a very late diagnosis compared to those diagnosed in the ward/inpatient unit. Having a higher education and university education were protective factors against having a very late diagnosis of HIV infection compared to elementary school education only. In addition, male patients were more likely to have a very late diagnosis compared to female patients. Conclusions: This study evidenced a high prevalence of late and very late diagnoses. Therefore, attention should be directed towards factors related to late and very late presentation.

Factors Associated with Late Diagnosis of Human Immunodeficiency Virus/Acquired Immunodeficiency Syndrome (HIV/AIDS) in a University Hospital in Brazil: Challenges to Achieving the 2030 Target

This study examined the prevalence of HIV late diagnosis (LD) and identified associated factors with LD among people living with HIV (PLWH). We extracted sociodemographic, epidemiological, and immunological information between 2018 and 2021 in Hunan, China from the HIV/AIDS Comprehensive Response Information Management System of China. The chi-squared test and multivariable logistic regression model were used to identify the factors associated with LD. Among 22,504 PLWH, 14,988 (66.6%) were diagnosed late. PLWH aged 50 and older had a higher proportion of LD (71.2%) than the younger group (60.0%). Older age, being male, Han ethnicity, being registered in Western Hunan, being transferred from health facilities, and being infected through heterosexual intercourse were associated with LD. Among PLWH younger than 50 years, apart from the factors mentioned above, individuals who had primary school or lower education, were non-student, and were divorced or widowed were more likely to be diagnosed late. Unlike younger PLWH, these factors were not associated with LD in the older group. But regional disparities in LD were more significant among them. LD in PLWH remains a severe issue, especially among older people. The study findings provided valuable insights into designing programs targeting groups at higher risk of LD to reduce its prevalence. 

Unveiling trends in late diagnosis among 22,504 people living with HIV in Hunan, China

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.

/pdf/real-world-data-in-late-presentation-of-hiv-infection-in-e3v1pwj9.pdf

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

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.

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

Objective
In order to investigate the epidemical characteristics of the Influenza-like illness (ILI) in Suzhou from 2010 to 2014, providing the scientific supports for influenza prevention and control of Suzhou.


Methods
The influenza samples were collected from the department of respiration of Suzhou Municipal Hospital Division and Suzhou city children's hospital, using fluorescent-PCR method to detect influenza virus nucleic acid and classifying influenza virus subtypes.


Results
8035 throat swabs of ILI specimens were collected from 2010-2014 in Suzhou, 959 samples were positive (11.94%), the positive rate of difference of adjacent years was statistically significant (χ2=77.78, P<0.0001). The positive rate was highest in the age group of 10-19 and lowest in the age group of 40-49 among all detected age groups. The influenza epidemic season were winter-spring and summer in Suzhou, type B influenza main in spring and winter, and influenza A in prevalent throughout all year, each subtype showing alternating popular.


Conclusions
From 2011-2014, influenza virus infected people in every age group of Suzhou, the peak popular time are winter-spring and summer of every year, all kinds of influenza subtype epidemic alternately, so it needs to supervise influenza etiology of Suzhou for a long-term to providing theoretical basis for further control the spread of the flu.


Key words: 
Influenza; Etiology; Surveillance

Survey of the epidemic characteristics of influenz in Suzhou from 2010 to 2014

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

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

Survey of the epidemic characteristics of influenz in Suzhou from 2010 to 2014