Molecular Biology Tests Research Articles

Cerebral thrombus analysis as a useful diagnostic tool for infective endocarditis in ischemic stroke patients.

Infective endocarditis (IE) is a life-threatening condition and a rare cause of ischemic stroke (IS). This study aimed to evaluate the utility of analyzing cerebral thrombi, obtained through endovascular thrombectomy in IS, for the pathological diagnosis of IE. Cerebral thrombi from three groups of IS patients were compared: definite IE (n = 10), cardioembolic stroke without and with concomitant infection (CE-I-: n = 30, CE-I+: n = 10). We performed histological examination, molecular biology, and microbiological tests on cerebral thrombi, to detect microorganisms and assess their composition. Median age of included patients was 73 years and 50% were females. Hematoxylin & Eosin and Grocott-Gomori Methenamine Silver stains detected microorganisms in all IE cerebral thrombi, and none in the control groups. Thrombus PCR detected relevant microorganism in n = 2/7 IE. Compared to control groups, IE thrombi were characterized by significant lower content of red blood cells (median [IQR]: IE = 7.4 [4.2-26.7], CE-I- = 49.3 [17-62.6], CE-I+ = 57.5 [40.7-60.8], % over thrombus section area [%TSA], p = 0.001), increased von Willebrand Factor (IE = 23.9 [19.1-32], CE-I- = 11.2 [8.2-12.8], CE-I+ = 12.9 [10.7-18.3], %TSA, p = 0.001), cell-dominant pattern of Neutrophil Extracellular Traps (IE = 100%, CE-I- = 69%, CE-I+ = 70%, p ⩽ 0.001), and more frequent sub-acute or chronic thrombus age classification (p ⩽ 0.001). These latter thrombus features displayed good discriminative ability between IE and controls, with AUC values between 0.84 and 0.95. Multimodal analysis of cerebral thrombi in IS with suspected IE supports early and definite pathological diagnosis by detecting pathogens and assessing changes in thrombus composition.

Zoonotic Bacteria and Vector-Borne Protozoa in Troglophilus Bat Colonies in Sicily (Southern Italy): A Biomolecular Survey.

Bats, as members of the order Chiroptera, are vital to ecosystems and serve as reservoirs for numerous microorganisms, some of which can cause zoonotic diseases. Human interactions with bats are increasing due to habitat alterations, making it essential to understand their microbiota, particularly potential pathogens. This study aimed to evaluate the excretion of zoonotic bacteria and protozoa in insectivorous bats from four caves in the provinces of Ragusa, Catania, and Syracuse (Sicily, Southern Italy) using molecular biology tests for zoonotic agents, including Bartonella henselae, Borrelia, Coxiella burnetii, Leptospira, Chlamydia, Rickettsia, Anaplasma, and Piroplasmids. From December 2020 to April 2023, urine, fecal swabs, ocular conjunctival swabs, and oropharyngeal swabs were collected from 149 bats of six species, along with guano samples from the caves. Bartonella henselae DNA was detected in 3 of the 149 tested bats, one ocular conjunctival swab and two oropharyngeal swabs. Chlamydia spp. DNA was detected in a sample of guano, in feces, ocular conjunctival and oropharyngeal swabs of a bat, and in four urine samples. Piroplasmid DNA was detected in 10 of 149 fecal swabs and in 5 of 16 bat ectoparasites. No samples were positive for Leptospira spp., Borrelia spp., Coxiella burnetii, Rickettsia spp., or Anaplasma spp. These findings underscore the importance of multiple sample types in assessing bats as reservoirs for zoonotic pathogens, particularly highlighting their role in transmitting pathogens through various body habitats, including saliva, urine, and ocular excretions. This study highlights the relevance of monitoring bat populations and studying their microbiota to enhance protections for both human and animal health.

MAPK14/AIFM2 pathway regulates mitophagy-dependent apoptosis to improve atrial fibrillation.

MAPK14/AIFM2 pathway regulates mitophagy-dependent apoptosis to improve atrial fibrillation.

Building a teaching method for “clinical molecular biology testing technology” guided by the cultivation of thinking skills

AbstractThe knowledge of “clinical molecular biology testing technology” is complex, conceptual expressions are abstract and difficult to understand, and the student's interest in learning is low. This study aimed to evaluate the effectiveness of a cyclic teaching method based on case analysis combined with an exploratory teaching method using mind mapping as an assignment. Students from the 2019 cohort of medical laboratory technology at Hunan University of Chinese Medicine served as the control group and received conventional lecture‐based teaching methods. Students from the 2021 cohort served as the test group and were undergoing teaching methods that focused on developing critical thinking skills. Course regular grades, exam scores, teaching evaluations, and questionnaires were used to evaluate the teaching effect. The test group had higher scores than the control group. It is also better in terms of memorization ability and comprehensive analytical ability. However, there were no differences between the two groups in terms of understanding and application ability. The test group showed higher satisfaction in improving knowledge mastery, cultivating learning interest, enhancing logical thinking ability, and utilizing post‐class assignments in the form of mind mapping for better knowledge consolidation and expansion than did the control group. The survey results from the test group indicate that a teaching method focused on developing thinking skills has significant effects on stimulating learning interest, improving cognitive ability, and helping to address clinical issues. Combining the cyclic teaching method based on case analysis with the exploratory teaching method using mind mapping as assignments can enhance learning motivation, facilitate knowledge acquisition, and is suitable for teaching in the discipline of “clinical molecular biology testing technology”.

MMLV reverse transcriptase, a relevant tool for molecular biology

The reverse transcriptase of Moloney Murine Leukemia Virus (MMLV) is an enzyme that synthesizes DNA from an RNA template. Among reverse transcriptases, this enzyme is currently the most commonly used in molecular biology and diagnostics. Since its discovery, this viral protein has been extensively studied, shedding light on its structural and functional characteristics, and offering opportunities to optimize the catalytic performances for biotechnological applications. The review describes the structural motifs of the reverse transcriptase of MMLV, the key amino acids in enzymatic catalysis and the enzyme's properties such as processivity, fidelity and thermal stability. This review reports the optimizations made to improve these parameters, which enabled the modified enzymes to be integrated in the molecular biology tests used in the laboratory on a daily basis.

Tetramethylpyrazine inhibits ferroptosis in spinal cord injury by regulating iron metabolism through the NRF2/ARE pathway.

Tetramethylpyrazine (TMP) is a natural alkaloid compound with antioxidant and neuroprotective effects. We hypothesized that TMP could exert neuroprotective effects by inhibiting ferroptosis through modulating iron metabolism, but its mechanism is unclear. Through in vivo and in vitro experiments, we have explored how TMP can regulate neurons' iron metabolism through the NRF2/ARE pathway to Inhibit ferroptosis. In the in vivo experiment, the effects of TMP on nerve function and secondary spinal cord injury were observed through behavioral tests and morphology staining. Transmission electron microscopy, molecular biology tests and immunofluorescence staining were used to investigate the role of TMP in the regulation of iron metabolism and ferroptosis through the Nrf2/ARE pathway. Using in vitro experiments to investigate the mechanism of TMP in inhibiting ferroptosis through the Nrf2/ARE pathway. Firstly, through in vivo experiments, we found that TMP improves motor function of rats with spinal cord injury, reduces spinal cord tissue damage and nerve cell death caused by secondary injury. Moreover, neuronal death and the formation of spinal cord cavities are inhibited by TMP. By regulating lipid peroxidation, TMP can inhibit mitochondrial damage and reduce ROS accumulation. Our study also demonstrated that TMP regulates iron metabolism through the NRF2/ARE pathway to inhibit ferroptosis and repair spinal cord injury. To further explore the regulatory mechanisms of TMP we down-regulating Nrf2 expression in subsequent in vitro experiments. We find that a key ferroptosis pathway, lipid peroxidation, can be regulated by TMP. Additionally, TMP inhibits iron overload-mediated ferroptosis by increasing Nrf2 transcriptional activity. A regulatory effect of TMP on the NRF2/ARE pathway was found in both in vitro and in vivo experiments. It promotes the transcription and translation of iron metabolizing and antioxidant molecules. Our study explored the inhibitory effect of TMP on ferroptosis from the iron metabolism pathway and provided new ideas for the treatment of SCI.

Coinfection of Mycoplasma suis and porcine circovirus type 3 is linked to reproductive failure in pig farms.

Reproductive disorders in swine herds pose significant challenges to pig breeding due to both infectious and non-infectious factors. In large-scale pig farming, coinfections are increasingly common, affecting sow health and herd productivity. This study aimed to determine occurrence and coinfection patterns of Mycoplasma suis and porcine circovirus type 3 in Vietnamese pig farms and to evaluate their association with reproductive disorders and clinical signs in affected herds. We collected 291 samples from 15 farms, composed of whole blood and various tissues from fetuses and weak-born piglets. Molecular biological testing was conducted to detect key pathogens of interest. Consistently, porcine circovirus type 3 (PCV3) and porcine Hemoplasma were detected and sequenced for the whole genome and partial 16S rRNA, respectively. The genetic diversity of PCV3 and Mycoplasma suis was analyzed. Various clinical signs, including abortion, stillborn, mummified, and weak-born piglets, and dermatitis, were recorded. M. suis was detected in 252/291 (86.59%) samples from all 15 surveyed farms, with an occurrence of 100%. PCV3 was detected in 35.05% (102/291) samples and 73.3% (11/15) of farms. PCV3 and M. suis coinfections were observed in 29.21% of the positive samples. It should be noted that most PCV3 Ct-values were above 30, indicating the existence of PCV3 in the herd but with insufficient data to confirm its pathogenic potential. The complete genomes of 10 PCV3 strains identified in this study exhibited high sequence homology, with >97% nucleotide identity. In addition, the eight partial 16S rRNA porcine Hemoplasma sequences shared absolute identity with M. suis isolates from pigs in China and Germany. This report on the occurrence of M. suis and PCV3 in pigs from farms with reproductive failure provides important insights into the expanding global distribution of these pathogens. Our findings warrant further investigations of the pathogenic potential and economic implications of M. suis and PCV3 in pigs with reproductive failure in Vietnam.

EXTRAHEPATIC MANIFESTATIONS IN PATIENTS WITH ACUTE HEPATITIS E – PAZARDZHIK, BULGARIA 2014 – 2022

BACKGROUND: Hepatitis E is a global health issue, only partially understood. Bulgarian record started in 2019 and data is not sufficient. AIM: This research aims to analyze extrahepatic manifestation of acute hepatitis E in patients with hepatitis E from Pazardzhik region, between 2014 – 2022. MATERIALS AND METHODS: The analysis includes 247 patients with acute hepatitis E, treated at the Department of Infectious Diseases of Pazardzhik Multiprofile Hospital for Active Treatment, Bulgaria between 2014 – 2022. The methodology includes clinical observation, laboratory tests and medical imaging. The diagnosis was established by serological /ELISA for anti-HEV IgM, IgG detection/and molecular-biological tests /RT-PCR for HEV RNA detection/. RESULTS: We observed extrahepatic manifestations in 19% (47/247) of the cases. In 60% (28/47) comorbidities were present, and 9% (4/47) were with underlying acute/chronic coinfection with another hepatotropic virus. Thrombocytopenia was found in 83% (39/47) of the patients; asymptomatic creatine kinase elevation – in 13% (6/47), acute pancreatitis – in 9% (4/47), transitory renal impairent – in 6% (3/47); 2% (1/47) had Guillain-Barré syndrome (GBS), 2% (1/47) – arrhythmia and 13% (6/47) – multiorgan involvement. While 91% (43/47) of the patients recovered, in 9% (4/47) the outcome was fatal. CONCLUSION: Extrahepatic manifestations might prevail, potentially delaying diagnosis of HEV-infection. Symptoms associated with comorbidities might also impede the final diagnosis. A diagnostic algorithm is needed to enhance the accurate diagnosis of HEV in patients with dubious symptoms.

Preanalytical Impact of Incomplete K2EDTA Blood Tube Filling in Molecular Biology Testing.

The aim of this study was to investigate the possible preanalytical effect of incomplete filling of blood tubes on molecular biology assays. The study population consisted of 13 healthy volunteers from whom 11 mL of whole blood was collected and then distributed in different volumes (1.5, 3.0, and 6.0 mL, respectively) into three 6.0 mL spray-dried and evacuated K2EDTA blood tubes. Automated RNA extraction was performed using the Maxwell® CSC RNA Blood Kit. DNA was extracted with a MagCorePlusII, with concomitant measurement of glyceralde-hyde-3-phosphate dehydrogenase (GAPDH) gene expression. The nucleic acid concentration was calculated using the NanoDrop 1000 spectrophotometer, and purity was assessed using A260/280 and A260/230 absorbance ratios. The RNA concentration was higher in the tubes filled with 1.5 and 3.0 mL of blood than in the reference 6 mL filled tube. The RNA 260/280 and RNA 260/230 ratios did not differ significantly between the differently filled blood tubes. The DNA concentration remained constant in the differently filled tubes. Compared to the 6.0 mL reference filled tube, the 1.5 mL and 3.0 mL filled blood tubes displayed a lower DNA 260/280 nm ratio. The DNA 260/230 ratio did not differ significantly in any of the variously filled tubes. Compared to the 6.0 mL reference filled blood tube, the 1.5 mL and 3.0 mL filled blood tubes showed a significant increase in the GAPDHcycle threshold. Our results suggest that underfilling of K2EDTA blood tubes may be a modest but analytically significant source of bias in molecular biology testing.

Research Progress in the Prevention and Control of Hospital-acquired Infections in Neonatal Units

Neonatal hospital-acquired infections are an important problem affecting the health and life safety of newborns. This paper reviews the research progress on the prevention and control of hospital-acquired infections in neonatology, discusses the risk factors of hospital-acquired infections in neonatology, summarizes the effective preventive and control measures such as strengthening the training and education on hospital-acquired infections, reducing invasive diagnostic and therapeutic operations, implementing disinfecting and isolating measures, rationally using antibiotics, and improving immunity for newborns, etc., and introduces common monitoring and detection techniques such as bacterial culture and drug susceptibility testing, molecular biology testing techniques, and biomarker detection and application. marker detection and application, and other commonly used surveillance and detection techniques. Future research should focus on strengthening the monitoring and study of pathogen resistance, optimizing the diagnosis and treatment process, strengthening the infection prevention and control training for healthcare workers, improving the infection prevention and control system in neonatal hospitals, and carrying out multidisciplinary collaborative research, so as to effectively reduce the incidence of infections and safeguard the health and safety of neonates.

Isolation of H2S-generating Bacterium (Desulfovibrio sp.) and Vibrio parahaemolyticus From Aquatic Farming and In Vitro Evaluation of the Ability of Bacteriophages as Biocontrol

Graphical Abstract Highlight Research H2S-generating bacterium (Desulfovibrio) and Vibrio sp. Were idenfied and surveyed its charecteristics. Bacteriophages, ɸTT1H, ɸTT2H, and ɸA2223, could reduce Desulfovibrio vulgaris and Vibrio parahaemolyticus colony sizes and change the bacterial shapes. The bacteriophages could not reduce Desulfovibrio vulgaris and Vibrio parahaemolyticus colony quantity. The bacteriophages affected neither the nucleotide sequence ToxR genes of Vibrio parahaemolyticus nor the 16S rRNA of Desulfovibrio vulgaris. Abstract Shrimp farming is an important industry in many countries. However, the leftover feed in shrimp ponds can create harmful compounds like H2S and provide a breeding ground for Vibrio bacteria, which causes acute hepatopancreatic necrosis disease. Antibiotics are commonly used to treat this disease, but they can lead to bacterial resistance and environmental pollution. Therefore, using bacteriophages as a treatment option is a more sustainable approach. The present study aimed to isolate H2S-generating bacteria and bacteriophages capable of inhibiting Vibrio sp. and Desulfovibrio sp. from shrimp pond water. Bacteria were identified through biochemical and molecular biology tests. The study utilized plaque and spread methods to observe changes in bacterial number and colony morphology. The study successfully isolated the bacterial strain Desulfovibrio vulgaris (12D) from shrimp ponds. Three potential bacteriophage strains, ɸTT1H, ɸTT2H, and ɸA2223, were identified that have the ability to inhibit Desulfovibrio vulgaris and V. parahaemolyticus bacteria by altering the size, shape, and number of colonies in treatments supplemented with phages. Although they do not alter the nucleotide sequence of these two bacterial strains, they still have a significant effect on controlling the bacterial population. Among the three potential bacteriophage lineages, ɸTT2H was able to inhibit Desulfovibrio vulgaris, reducing the colony quantity by 2.9%. This research allowed researchers to apply bacteriophages to shrimp culture.

Parotid Gland Tumors: Molecular Diagnostic Approaches.

Parotid gland pathology represents a web of differential diagnoses. There are many complex cases that require extensive diagnostic tests for a complete and correct final pathology diagnosis. Currently the official classification of parotid gland tumors extends over more than 40 subtypes. We performed a query of the PubMed database regarding the use of molecular biology tests in performing a better characterization of the tumors in specific cases. By using fluorescence in situ hybridization (FISH), reverse transcription polymerase chain reaction (RT-PCR) or next-generation sequencing, the team managing complex cases can offer a personalized therapeutic solution. We review the molecular differential diagnosis according to published articles in the last 5 years for many types of parotid gland tumors ranging from benign to borderline malign tumors to malign aggressive tumors. Mucoepidermoid carcinoma is a distinct subtype of parotid malignancy that was the subject of a consistent number of articles. However, the molecular biology diagnosis techniques helped more in excluding the diagnosis of mucoepidermoid carcinoma, and probably retrospectively limiting the number of cases with this final diagnosis. In Romania, the molecular biology diagnosis is available only in limited research facilities and should receive more consistent funding that will make it available on a larger scale. The novelty of this scoping review is that we propose an algorithm for molecular differential diagnosis of the tumors that could be encountered in the parotid gland.

Surpassing the natural limits of serological diagnostic tests

Surpassing the natural limits of serological diagnostic tests

Evaluation of changes caused by HR-HPV infection in squamous cell carcinoma of the head and neck using Raman microspectroscopy in combination with multivariate statistical analysis

Abstract Introduction: Squamous cell carcinoma of the head and neck region (HNSCC), with a positive status for high oncogenic potential human papillomavirus (HR-HPV), represents a clinically distinct disease entity compared to HPV-independent cases. Patients exhibit variations in prognosis and proposed therapy regimens. A prompt and reliable diagnosis of the presence of HPV infection could optimize the treatment for these patients. Currently employed treatment methods are long-term, expensive, and lack specificity, especially when administered separately. Material and methods: The research objective of this study is to explore significant differences in the Raman spectra of biological samples taken from patients with HNSCC, facilitating the confirmation of HPV virus presence. Study groups were delineated based on histopathological diagnosis and molecular biology tests, confirming the biological activity of the virus and the presence of the HR-HPV form with a diagnosis of a specific subtype. Results: To identify high oncogenic potential human papillomavirus (HR-HPV) infection as a crucial factor in squamous cell carcinoma of the head and neck region, an effective automatic data analysis system was established, relying on Raman microspectroscopy and multivariate analysis. Our results showed clear ranges of the Raman spectrum that differentiated between HPV-associated and non-HPV-associated cancers. Conclusions: In conclusion, our experience shows a great diagnostic potential of Raman confocal microscopy with multidimensional statistical analysis. In the future, the use of this method may allow for the creation of an effective and automated HR-HPV detection system in neoplastic tissue.

"Primum, non nocere": The Epidemiology of Toxigenic Clostridioides difficile Strains in the Antibiotic Era-Insights from a Prospective Study at a Regional Infectious Diseases Hospital in Eastern Europe.

Clostridioides difficile infection (CDI), though identified nearly five decades ago, still remains a major challenge, being associated with significant mortality rates. The strains classified as hypervirulent, notably 027/NAP1/BI, have garnered substantial attention from researchers and clinicians due to their direct correlation with the severity of the disease. Our study aims to elucidate the significance of toxigenic Clostridioides difficile (CD) strains in the clinical and therapeutic aspects of managing patients diagnosed with CDI. We conducted a single-center prospective study, including patients with CDI from north-eastern Romania. We subsequently conducted molecular biology testing to ascertain the prevalence of the presumptive 027/NAP1/BI strain within aforementioned geographic region. The patients were systematically compared and assessed both clinically and biologically, employing standardized and comparative methodologies. The study enrolled fifty patients with CDI admitted between January 2020 and June 2020. Among the investigated patients, 43 (86%) exhibited infection with toxigenic CD strains positive for toxin B genes (tcdB), binary toxin genes (cdtA and cdtB), and deletion 117 in regulatory genes (tcdC), while the remaining 7 (14%) tested negative for binary toxin genes (cdtA and cdtB) and deletion 117 in tcdC. The presence of the presumptive 027/NAP1/BI strains was linked to a higher recurrence rate (35.56%, p = 0.025), cardiovascular comorbidities (65.1% vs. 14.2%, p = 0.016), and vancomycin treatment (55.8% vs. 14.3%, p = 0.049). The findings of our investigation revealed an elevated incidence of colitis attributed to presumptive 027/NAP1/BI. Despite the prevalence of the presumptive 027 strain and its associated heightened inflammation among the patients studied, no significant differences were observed regarding the clinical course or mortality outcomes.

Introduction of a decalcification method for bone marrow biopsy tissue

Bone marrow biopsy is one of the important means of hematopathological diagnosis, which has decisive diagnostic significance for various benign and malignant lymphohematopoietic system diseases. Its diagnostic value includes morphological observation, immunohistochemistry, genetics, and molecular biology testing. Owing to the unique nature of bone marrow biopsy, decalcification is an essential step in the pre-treatment process. Its purpose is to remove calcium from bone tissue, preserve intact collagen fiber components, facilitate tissue sectioning, and prevent tissue detachment during staining. If bone marrow biopsy lacks sufficient decalcification, preparing a section is difficult. Conversely, if decalcification is excessive, it can seriously disrupt tissue antigen activity. Therefore, a decalcification method with high decalcification efficiency and mild antigen damage is essential for bone marrow biopsy. This article introduces a bone marrow biopsy tissue decalcification method with high efficiency and less antigen loss: decalcification is performed at room temperature with 12% formic acid and 8% hydrochloric acid decalcification solution on a shaker.

Analysis of different detection methods for pulmonary tuberculosis

Tuberculosis is classified as a chronic respiratory disease with its pathogen being Mycobacterium tuberculosis (MTB). It poses a significant health threat due to its highly contagious nature and is difficulty in treatment. Various methods are employed to test for tuberculosis, including pathological, immunological and molecular biological tests. Each test has its unique principles and all are different in effects, and are widely used in clinical practice. In this research, principles, development and effects of each type of tests in testing pulmonary tuberculosis will be explored respectively. Additionally, the review will provide insights into the optimization of tuberculosis testing by comparing and contrasting different testing methods. Pathological tests represent the most traditional approach to testing for tuberculosis. Bacterial culture, a pathological test, is the gold standard of tuberculosis test. Nevertheless, it suffers from a long detection time. Meanwhile, immunological tests and molecular biological tests benefits from a fast detection, while the accuracy of the tests can be influenced by multiple factors. Thus, it will accelerate the detection speed while enhancing the precision to combine three different kinds of tests in clinical practice.

Etiological diagnostic methods and research progress of forest encephalitis

Forest encephalitis is a natural focal disease transmitted through the bite of hard ticks, and its pathogen is the tick-borne encephalitis virus from the Flaviviridae family. The mortality rate of forest encephalitis is relatively high, making laboratory testing significant in diagnosing this disease. This article elaborates on the etiological diagnostic methods and recent research progress in forest encephalitis. Laboratory tests for forest encephalitis mainly include routine examinations, serological tests, virus isolation, and molecular biological testing. The detection of serum-specific IgM antibodies against the forest encephalitis virus is of great importance for early diagnosis, and specific IgG antibodies serve as a "gold standard" for differentiation from other diseases. Techniques such as enzyme-linked immunosorbent assay (ELISA) or indirect immunofluorescence assay for detecting specific IgM antibodies in serum and/or cerebrospinal fluid, the serum hemagglutination inhibition test or serum complement fixation test, and the double serum hemagglutination inhibition test or complement fixation test all contribute to the early diagnosis. The development of molecular testing methods is rapid, and techniques such as metabolomics, digital PCR, and matrix metalloproteinases are also applied in the early diagnosis of forest encephalitis.

Hepatitis E outbreak in the health district of Bocaranga-Koui, Central African Republic, 2018–2019

BackgroundHepatitis E virus (HEV) is a major public health disease causing large outbreaks and sporadic cases of acute hepatitis. We investigated an outbreak of HEV infection that occurred in September 2018 in the health district (HD) of Bocaranga-Koui, located in the northwestern part of Central African Republic (CAR).MethodsBlood samples were collected from 352 patients aged 0–85 years suspected to be infected with yellow fever (YF), according to the World Health Organization YF case definition. The notification forms from recorded cases were used. Water consumed in the HD were also collected. Human samples found negative for anti-YF IgM were then tested by ELISA for anti-HEV IgM and IgG antibodies. Positive anti-HEV (IgM and/or IgG) samples and collected water were then subjected to molecular biology tests using a real time RT-PCR assay, followed by a nested RT-PCR assay for sequencing and phylogenetic analysis.ResultsOf the 352 icterus patients included, anti-HEV IgM was found in 142 people (40.3%) and anti-HEV IgG in 175 (49.7%). Although HEV infection was detected in all age groups, there was a significant difference between the 0–10 age groups and others age groups (P = 0.001). Elevated levels of serum aminotransferase were observed in anti-HEV IgM-positive subjects. Phylogenetic analysis showed HEV genotype 1e in infected patients as well as in the contaminated water.ConclusionThis epidemic showed that CAR remains an HEV-endemic area. The genotype 1e strain was responsible for the HEV outbreak in Bocaranga-Koui HD. It is necessary to implement basic conditions of hygiene and sanitation to prevent further outbreaks of a HEV epidemics, to facilitate access to clean drinking water for the population, to launch intensive health education for basic hygiene measures, to sett up targeted hygiene promotion activities and, finally, to ensure that formal health care is available.

Rapid detection of fish with SVC symptoms based on machine vision combined with a NAM-YOLO v7 hybrid model

Rapid detection of fish with SVC symptoms based on machine vision combined with a NAM-YOLO v7 hybrid model