Data_Sheet_1_A novel detection method for the pathogenic Aeromonas hydrophila expressing aerA gene and/or hlyA gene based on dualplex RAA and CRISPR/Cas12a.PDF

Aeromonas hydrophila is an emerging waterborne and foodborne pathogen with pathogenicity to humans and warm water fishes, which severely threatens human health, food safety and aquaculture. A novel method for the rapid, accurate, and sensitive detection of pathogenic A. hydrophila is still needed to reduce the impact on human health and aquaculture. In this work, we developed a rapid, accurate, sensitive, and visual detection method (dRAA-CRISPR/Cas12a), without elaborate instruments, integrating the dualplex recombinase-assisted amplification (dRAA) assay and CRISPR/Cas12a system to detect pathogenic A. hydrophila expressing aerA and/or hlyA virulence genes. The dRAA-CRISPR/Cas12a method has high sensitivity, which can rapidly detect (about 45 min) A. hydrophila with the limit of detection in 2 copies of genomic DNA per reaction, and has high specificity for three pathogenic A. hydrophila strains (aerA+hlyA−, aerA−hlyA+, and aerA+hlyA+). Moreover, dRAA-CRISPR/Cas12a method shows satisfactory practicability in the analysis of the spiked human blood and stool and fish samples. These results demonstrate that our developed pathogenic A. hydrophila detection method, dRAA-CRISPR/Cas12a, is a promising potential method for the early diagnosis of human A. hydrophila infection and on-site detection of A. hydrophila in food and aquaculture.</p

Genotype profiles and co-infection of HAdV.

<p>*, indicated the statistically difference of the HAdV infection rate between the Northern and Eastern China.</p><p>Genotype profiles and co-infection of HAdV.</p

Demographic data in this study.

<p>Demographic data in this study.</p

Molecular Typing and Epidemiology Profiles of Human Adenovirus Infection among Paediatric Patients with Severe Acute Respiratory Infection in China

<div><p>Background</p><p>Human adenoviruses (HAdVs) have been recognised as pathogens that cause a broad spectrum of diseases. The studies on HAdV infection among children with severe acute respiratory infection (SARI) are limited.</p><p>Objective</p><p>To investigate the prevalence, epidemiology, and genotype of HAdV among children with SARI in China.</p><p>Study Design</p><p>Nasopharyngeal aspirates (NPAs) or induced sputum (IS) was collected from hospitalised children with SARIs in Beijing (representing Northern China; n = 259) and Zhejiang Province (representing Eastern China; n = 293) from 2007 to 2010. The prevalence of HAdV was screened by polymerase chain reaction (PCR), followed by sequence typing of PCR fragments that targeted the second half of the hexon gene. In addition, co-infection with other human respiratory viruses, related epidemiological profiles and clinical presentations were investigated.</p><p>Results and Conclusions</p><p>In total, 76 (13.8%) of 552 SARI patients were positive for HAdV, and the infection rates of HAdV in Northern and Eastern China were 20.1% (n = 52) and 8.2% (n = 24), respectively. HAdV co-infection with other respiratory viruses was frequent (infection rates: Northern China, 90.4%; Eastern China, 70.8%). The peak seasons for HAdV-B infection was winter and spring. Additionally, members of multiple species (Human mastadenovirus B, C, D and E) were circulating among paediatric patients with SARI, of which HAdV-B (34/52; 65.4%) and HAdV-C (20/24, 83.3%) were the most predominant in Northern and Eastern China, respectively. These findings provide a benchmark for future epidemiology and prevention strategies for HAdV.</p></div

Clustering analysis of genotyping data of B. cereus clinical endophthalmitis isolates.

<p>Phylogenetic tree of <i>B</i>. <i>cereus</i> clinical endophthalmitis isolates was drawn based on <i>vrrA</i> gene sequence analyses. The figure showed that <i>B</i>. <i>cereus</i> isolats could be grouped into three genotyping (GT) groups: GTI, GTII, and GTIII. GTI was be further divided into three subgroups.</p

Phylogenetic analysis of HAdV based on the partial hexon gene.

<p>The phylogenetic tree was constructed by the neighbour-joining method, and bootstrap values were determined by 1000 replications in MEGA5.0. (Prefix-N: samples from Northern China; Prefix-E: samples from Eastern China; ■, sequences of the reference strains of HAdV-A cut from genomes found in GenBank; ●, sequences of reference strains of HAdV-B; ▼, sequences of reference strains of HAdV-C; Δ, sequences of reference strains of HAdV-D; ○, sequence of reference strain of HAdV-E; ◆, sequences of reference strains of HAdV-F.</p

Seasonal distribution of HAdV infection.

<p>*, also contain one HAdV-D (HAdV-37) and one HAdV-E (HAdV-4)</p><p>**, indicated the statistically difference of the HAdV infection rate among four seasons</p><p>Seasonal distribution of HAdV infection.</p

Clinical manifestation and prognosis associated with three <i>vrrA</i>-based genotypes of <i>B</i>. <i>cereus</i>.

<p>Clinical manifestation and prognosis associated with three <i>vrrA</i>-based genotypes of <i>B</i>. <i>cereus</i>.</p

Age distribution of HAdV infection.

<p>*, also contain one HAdV-D (HAdV-37)</p><p>* *, also contain one HAdV-E (HAdV-4)</p><p>*** indicated the statistically difference of the HAdV infection rate among four age groups</p><p>Age distribution of HAdV infection.</p

B-scan images of the patients' eyes infected with <i>B</i>. <i>cereus</i> before (A,B,C) and after (D,E,F) surgery operation.

<p>(A). Infection caused by GTI strains showed severe vitreous opacity (gay arrows). (B). Infection caused by GTII strains showed mild vitreous opacity (gray arrows). (C). Mild vitreous opacity infection caused by GTIII strains (gray arrow),high reflection and ascoustic shadow in vitreous showed foreign body (white arrow). (D). Infection caused by GTI strains after surgery. (E). Infection caused by GTII strains after surgery. (F). Infection caused by GTIII strains after surgery(the false expansion of eye ball after vitreous surgery with silicon oil tamponade).</p