Listeria monocytogenes is widely present in the natural environment. This bacteria can cause infections in both humans and animals. In humans, the most vulnerable groups to be infected with L. monocytogenes are the elderly, people with an impaired immune system and chronically illness, pregnant women, and newborn babies. The aim of this study was to develop a multiplex PCR assay for the rapid detection of L. monocytogenes in mock clinical samples. A pair of primers were designed for detection of L. monocytogenes based on prs, a Listeria genus specific gene, and hly, a hemolysin gene. The specificity of the primers were tested by using different L. monocytogenes strains and other common pathogenic bacteria. The results showed that L. monocytogenes strains were positive in the detection and other tested strains were negative in mock (spiked) clinical samples. The sensitivity of multiplex PCR assay was 102 CFU/ml per reaction. The specificity and sensitivity of multiplex PCR technology for detecting L. monocytogenes in mock (spiked) clinical samples were high, and the assay could be completed within 1.5 hours. Therefore, this established multiplex PCR provides a rapid and reliable method and will be useful for the detection of L. monocytogenes in mock clinical samples.
Introduction
Leptospirosis is a neglected disease in Vietnam. Until now, there has been limited knowledge about risk factors of this disease in Vietnam. The study was carried out to identify agricultural and behavioral factors associated with the transmission of leptospirosis in Vietnam.
Methods
This matched retrospective hospital-community-based case–control study was conducted from 1 October 2018 to 31 October 2019. We recruited cases from 11 selected government hospitals in three provinces of Vietnam, while controls were selected from the same communes of cases and matched by age (± 2 years) and sex. Microscopic agglutination test (MAT) and enzyme-linked immunosorbent assay (ELISA) were applied to determine confirmed cases, while only MAT was used to identify controls with a single high MAT titer < 1:100.
Results
504 participants (252 cases and 252 controls) were identified. Cultivating (OR 2.83, CI 1.38–5.79), animal farming (OR 8.26, CI 2.24–30.52), pig owners (OR 10.48, CI 5.05–21.73), cat owners (OR 2.62, CI 1.49–4.61) and drinking unboiled water (OR 1.72, CI 1.14 –2.59, p = 0.010) were significantly associated with human leptospirosis in Vietnam. Hand washing after farming/ gardening (OR 0.57, CI 0.38–0.86, p = 0.007) and bathing after farming, gardening, contact with cattle and poultry (OR 0.33, CI 0.19–0.58, p = 0.000) were determined as protective factors for this disease.
Conclusions
In short, the case–control study has revealed the risks in agricultural and animal practices and protective behavioral factors related to human leptospirosis in Vietnam. The findings suggested promotion of communication and health education programs targeting health behaviors in daily life and agricultural practices. Using personal protective equipment such as gowns, gloves, and boots during agricultural practices, especially cultivating and animal farming, is most recommended.
Mục tiêu của nghiên cứu này là nhằm đánh giá sự khác nhau và hiệu quả của phương pháp IFA và PCR giải trình tự trong việc xác định sự hiện diện và định danh ở mức độ loài của Cryptosporidium spp. trong mẫu chất thải biogas. Trong nghiên cứu này, 167 mẫu chất thải biogas được thu thập từ 03 huyện (Việt Yên, Lạng Giang và Hiệp Hòa) của tỉnh Bắc Giang từ năm 2019 - 2020 để tìm bào nang Cryptosporidium. Có 16 trong số 167 (9,6%) mẫu được phân tích bằng phương pháp miễn dịch huỳnh quang (IFA) dương tính với Cryptosporidium spp.,. Trong số này 11 mẫu đồng thời được khẳng định dương tính với Cryptosporidium spp., bằng phương pháp PCR. Các mẫu dương tính với PCR được giải trình tự trên gen 18S rRNA. Cụ thể, có 4 loài Cryptosporidium scrofarum, 2 C. suis được phát hiện trong mẫu nước thải trước biogas. Có 2 loài C. scrofarum, 1 C. suis, 1 C. bailey và 1 C. meleagridis được phát hiện trong mẫu nước thải sau biogas. Kết quả PCR giải trình tự cho thấy các loài Cryptosporidium phân lập được là những loài lây truyền từ động vật sang người. Chính vì vậy, việc xác định Cryptosporidium spp. bằng phương pháp miễn dịch huỳnh quang, kết hợp với PCR giải trình tự gen 18S rRNA có ý nghĩa quan trọng trong các nghiên cứu dịch tễ học, điều tra nguồn gốc của nhiễm trùng và giám sát bệnh tiêu chảy do Cryptosporidium.
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