Cryptosporidium is an important intestinal parasite that is mainly transmitted through the fecal-oral route. Human infection may occur following ingestion of water and food contaminated by Cryptosporidium oocysts, and children and immunocompromised individuals are at a high risk of infections. The main symptoms of Cryptosporidium infections include diarrhea, vomiting, malnutrition, and even death. Because of high sensitivity and rapid procedures, molecular tests are helpful for the diagnosis of cryptosporidiosis and may reduce the public health risk of cryptosporidiosis. This review summarizes the advances in the latest prevalence and molecular detection of human Cryptosporidium infections during recent years.
［摘要］ 隐孢子虫是一种重要的肠道寄生虫, 主要经粪-口途径传播, 人摄入被隐孢子虫卵囊污染的水源和食物均有感 染风险。儿童及免疫力低下者为隐孢子虫感染高危人群, 感染后可以引起腹泻、呕吐、营养不良等, 严重者甚至导致死 亡。分子检测技术因反应快速、灵敏度高, 已广泛应用于隐孢子虫检测。本文对近年来人群隐孢子虫感染流行情况, 以 及隐孢子虫分子检测相关研究进展进行综述。.

SARS-CoV-2 is the agent responsible for the global pandemic sickness, COVID-19. It is an enveloped virus that belongs to the family Coronaviridae. Recent studies have revealed the fecal shedding of the virus and have been found to enter wastewater and aquatic systems. Prolonged viral presence in fecal samples is a common observation in the reported literature. Survival of the virus in the recipient environment could be a crucial factor that influences its fecal-oral transmission. The detection of a novel coronavirus in wastewater opportunity has potential for environmental surveillance at the community or population level. Such a surveillance system can enable the early detection of disease outbreaks in zones with pre-symptomatic/asymptomatic patients and act as a complementary tool for continuous monitoring of quarantine zones. In contrast to developed regions, resource constraints in underdeveloped communities coupled with different sanitation settings may pose a challenge to wastewater sampling and surveillance. To begin, this review summarizes the literature on the presence of SARS-CoV-2 in feces. The approaches for viral extraction, concentration, and detection in wastewater matrices are then highlighted. Finally, investigations on wastewater-based epidemiology for SARS-CoV-2 surveillance are reviewed.

Rapid and accurate detection technologies are crucial for disease prevention and control. In particular, the COVID-19 pandemic has posed a great threat to our society, highlighting the importance of rapid and highly sensitive detection techniques. In recent years, CRISPR/Cas-based gene editing technique has brought revolutionary advances in biotechnology. Due to its fast, accurate, sensitive, and cost-effective characteristics, the CRISPR-based nucleic acid detection technology is revolutionizing molecular diagnosis. CRISPR-based diagnostics has been applied in many fields, such as detection of infectious diseases, genetic diseases, cancer mutation, and food safety. This review summarized the advances in CRISPR-based nucleic acid detection systems and its applications. Perspectives on intelligent diagnostics with CRISPR-based nucleic acid detection and artificial intelligence were also provided.

Pythiosis, a life-threatening infectious condition caused by Pythium insidiosum, has been increasingly reported in humans and animals worldwide. Antifungal drugs usually fail to control the pathogen. The surgical removal of an infected organ is the treatment of choice. Many affected patients die due to advanced infection. A timely and accurate diagnosis could lead to a better prognosis in pythiosis patients and save their lives. Although a standard culture method is available in microbiological laboratories, it is time-consuming, laborious, and insensitive for P. insidiosum identification. Immunological assays have been developed to improve the diagnosis of pythiosis. However, immunological methods are commercially unavailable and primarily detect anti-P. insidiosum antibodies, which constitute indirect evidence of pythiosis, making it challenging to differentiate a past from a recent infection. Moreover, such immunological tests cannot diagnose patients with a local infection, such as in the eye. Nucleic acid-based tests (NATs) are efficient for the direct and rapid detection of P. insidiosum DNA in trace-amount or culture-negative specimens. The reagents and equipment required for NATs are usually available in molecular diagnostic laboratories. Herein, we provide a systematic review to comprehensively present the principal and clinical usages, advantages, and limitations of such NATs in the detection of P. insidiosum. Various NATs have been established to detect P. insidiosum, which can be classified into amplification-based (i.e., PCR assays, isothermal tests, and next-generation sequencing methods) and non-amplification-based (i.e., DNA hybridization) techniques. This concise review on NATs constitutes an up-to-date reference with which healthcare professionals can learn about and decide upon which detection method is suitable for their respective laboratory environments.

Biosensors play a central role in moving diagnostics to being on-site or decentralized. Affinity biosensor, an important category of biosensors, has important applications in clinical diagnosis, pharmaceuticals, immunology, and other fields. Affinity biosensors rely on specific binding between target analytes and biological ligands such as antibodies, nucleic acids, or other receptors to generate measurable signals. Oftentimes the target analytes in practical samples are of low abundance in a complex matrix. Traditional affinity biosensors mainly rely on random diffusion of analytes in solution to conjugate with biorecognition elements on the sensor surface of electrodes. The process may take hours or even days, which is not conducive to rapid and sensitive detection of biosensors. Therefore, it is strongly desired to incorporate an enrichment mechanism for target analytes into biosensor-based detection. AC electrokinetic (ACEK) effect can realize rapid enrichment of analytes by application of AC electric fields, which holds great promise for achieving high sensitivity, low detection limit, and rapid turnaround. This article reviews the studies of affinity biosensors integrated with ACEK enrichment in the past decade, and summarizes the latest detection methods, detection devices and applications, hoping to provide some insights and references for researchers in related fields.

Worldwide, Mycoplasma gallisepticum (MG) and M. synoviae (MS) are the main agents responsible for chronic respiratory disease in poultry. Therefore, we conducted a systematic review and meta-analysis to estimate their occurrence. We searched electronic databases to find peer-reviewed publications reporting the molecular detection of MG and MS in poultry and used meta-analysis to estimate their pooled global occurrence (combined flock and individual), aggregating results at the regional and national levels. We performed a subgroup meta-analysis for subpopulations (broilers, layers, breeders and diverse poultry including turkeys, ducks and ostriches) and used meta-regression with categorical modifiers. We retrieved 2294 publications from six electronic databases and included 85 publications from 33 countries that reported 62 studies with 22,162 samples for MG and 48 studies with 26,413 samples for MS. The pooled global occurrence was 38.4% (95% CI: 23.5-54.5) for MS and 27.0% (20.4-34.2) for MG. Among regions, Europe and Central Asia had the lowest occurrence for both pathogens, while MG and MS were highly prevalent in South Asia and sub-Saharan Africa, respectively. At the national level, MG occurrence was higher in Algeria, Saudi Arabia and Sudan, whereas China, Egypt and Ethiopia reported higher values of MS. Among the poultry subpopulations, MS and MG were more prevalent in the breeders and layers (62.6% and 31.2%, respectively) than in diverse poultry. The year of publication, the sample size and the level of ambient air pollution (measured indirectly by PM2.5) were associated with the occurrence of both mycoplasmas. Our study revealed high and heterogeneous occurrence values of MG and MS and justifies the need for early detection and improved control measures to reduce the spread of these pathogens.

Cryptosporidium spp. is one of the prime agents of infectious diarrhea. Cryptosporidium spp. has been gaining awareness as a pathogen of public health importance in India and other developing countries. Owing to the nature of multiple transmission routes such as person-to-person, animal-to-person, waterborne and foodborne, the epidemiology of cryptosporidiosis in humans is not well known. A deeper understanding of the pathogenesis may lead to better diagnosis and better treatment of the condition. Asymptomatic human and animal transmission illustrates that the spread of infection through the environment is a more plausible explanation, waterborne transmission in particular. The disease burden is underestimated and its global impact is yet to be quantified due to the lack of country-specific estimates. Assessment of the disease itself has been crucial since the morphological indistinguishability, differences in distribution and transmission, and variations in the genotypes.

The zoonosis Q fever is caused by the obligate intracellular bacterium Coxiella burnetii. Besides the main transmission route via inhalation of contaminated aerosols, ticks are discussed as vectors since the first isolation of the pathogen from a Dermacentor andersonii tick. The rare detection of C. burnetii in ticks and the difficult differentiation of C. burnetii from Coxiella-like endosymbionts (CLEs) are questioning the relevance of ticks in the epidemiology of Q fever. In this review, literature databases were systematically searched for recent prevalence studies concerning C. burnetii in ticks in Europe and experimental studies evaluating the vector competence of tick species. A total of 72 prevalence studies were included and evaluated regarding DNA detection methods and collection methods, country, and tested tick species. Specimens of more than 25 different tick species were collected in 23 European countries. Overall, an average prevalence of 4.8% was determined. However, in half of the studies, no Coxiella-DNA was detected. In Southern European countries, a significantly higher prevalence was observed, possibly related to the abundance of different tick species here, namely Hyalomma spp. and Rhipicephalus spp. In comparison, a similar proportion of studies used ticks sampled by flagging and dragging or tick collection from animals, under 30% of the total tick samples derived from the latter. There was no significant difference in the various target genes used for the molecular test. In most of the studies, no distinction was made between C. burnetii and CLEs. The application of specific detection methods and the confirmation of positive results are crucial to determine the role of ticks in Q fever transmission. Only two studies were available, which assessed the vector competence of ticks for C. burnetii in the last 20 years, demonstrating the need for further research.

Detecting free tumor cells in the peritoneal lavage fluid of gastric cancer patients permits to assess a more accurate prognosis, predict peritoneal recurrence and select cases for a more aggressive treatment. Currently, cytology and molecular biology comprise the two most popular methods of detection that are under constant study by researchers.
We burrowed into the available literature comparing cytological with molecular detection of free intraperitoneal gastric cancer cells. PubMed, Science Direct, Scopus and Google Scholar were the search engines investigated.
As of 2017, 51 dedicated studies have been published. Messenger RNA of carcinoembryonic antigen was the genetic target most frequently described. The genetic technique is usually superior to cytology in sensitivity (38-100% vs. 12.3-67% respectively), whereas cytological examination tends to show a slight pre-eminence in specificity (approximately 100%).
So far, given the imperfection of each method, employment of both cytology and molecular examination seem to be mandatory.

Enterobacteriaceae are a common source of both community- and hospital-acquired infections. The resistance of Enterobacteriaceae to β-lactam antibiotics is a major public health problem and rapid detection is essential for the proper management of infections. Not only for the initiation of the proper antimicrobial regimen but also to stop the spread of multi-resistant bacteria. In this review, molecular methods that detect extended-spectrum-β-lactamase-harboring and carbapenem-resistant Enterobacteriaceae were discussed. More specifically the performances of assays that were tested in a clinical situation were explored. When compared with conventional phenotypical methods (the gold standard) all molecular methods generated results faster and showed a higher sensitivity. Because molecular methods only detect selected genes, it is important to have good knowledge on the local gene pool. Furthermore, result need to be interpreted within a defined epidemiological context. Therefore, at this moment molecular methods should be seen as an important tool to complement rather than replace conventional methods.