Development and Application of Real-Time PCR Assay for Detection of Salmonella Abortusequi

Wang, Jinhui; Guo, Kui; Li, Shuaijie; Liu, Diqiu; Chu, Xiaoyu; Wang, Yaoxin; Guo, Wei; Du, Cheng; Wang, Xiaojun; Hu, Zhe

doi:10.1128/jcm.01375-22

Cited by 9 publications

(5 citation statements)

References 31 publications

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“…Equine abortus salmonellosis has occurred frequently in recent years and has caused serious economic losses to the equine industry in China [4,5]. Terefore, the development of reliable serum diagnostic methods is essential to achieve the most efcient S. Abortusequi antibody detection.…”

Section: Discussionmentioning

confidence: 99%

“…Te methods for isolation and identifcation of S. Abortusequi were performed as our previously study described [5]. Briefy, vaginal swab liquid was plated on chromogenic Salmonella agar medium.…”

Section: Isolation and Identifcation Of S Abortusequimentioning

confidence: 99%

“…Salmonellosis is one of the most important bacterial diseases in several animals and has caused huge economic losses in the pig [1], poultry [2], pigeon [3], and equine industries [4,5]. Salmonella enterica subsp.…”

Section: Introductionmentioning

confidence: 99%

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Development and Application of an iELISA for the Detection of Antibody against Salmonella Abortusequi

Guo

Zhang

Yang

et al. 2023

Transboundary and Emerging Diseases

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Equine abortus salmonellosis is a bacterial disease that causes high abortion rates in susceptible equids and therefore significant economic losses. Although the tube agglutination test (TAT) is a commonly used serological test for S. Abortusequi, it is not highly specific or sensitive, and the development of more sensitive, specific and rapid assays is therefore urgently required. In this study, an indirect enzyme-linked immunosorbent assay (iELISA) was developed for the specific detection of flagellum protein (FljB) antibodies against S. Abortusequi. Negative sera from horses in China (n = 1030) were used to establish the baseline for a negative population, and reference antisera positive against other viruses or bacteria were used to test the cross reactivity of the technique. The performance of the FljB iELISA was evaluated against that of the standard TAT, and was tested using field serum samples. The FljB iELISA assay was 8–16 times more sensitive than TAT. ROC analysis showed that the FljB iELISA was accurate, with an area under the curve (AUC) = 0.9943 (95% CI, 0.9815–1.000). The diagnostic sensitivity (DSe) of the FljB iELISA was 98.9% (95% Cl, 93.84%–100.00%), which was higher than that of TAT (DSe 38.6; 95% CI, 29.14%–49.08%). The diagnostic specificity (DSp) of the iELISA was 100.0% (95% CI, 95.82%–100.0%). When the 508 clinical samples were tested, the FljB iELISA had a positive detection rate of 51.38% (261/508, 95% CI, 51.24%–51.51%), which was higher than that of TAT (44/508). We also performed a serological survey for S. Abortusequi infection, using a series of samples collected from across eighteen provinces of China in 2021. The results showed that all provinces except Jiangsu had a certain number of cases, and the positive rates ranged from 0% to 96.9%, indicating the wide spread of S. Abortusequi in China. The abovementioned results suggest that the FljB iELISA developed in this study is rapid, sensitive, specific, and repeatable and is likely to be a suitable test for large-scale serological surveys for the detection and control of S. Abortusequi infection.

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Section: Discussionmentioning

confidence: 99%

Section: Isolation and Identifcation Of S Abortusequimentioning

confidence: 99%

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Development and Application of an iELISA for the Detection of Antibody against Salmonella Abortusequi

Guo

Zhang

Yang

et al. 2023

Transboundary and Emerging Diseases

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“…The absence of specific clinical infections requires specific laboratory investigations. Furthermore, given often prolonged bacteriological analyses, several molecular methods have been developed, such as real-time polymerase chain reaction (PCR), and immunochemical methods such as enzyme-linked immunosorbent assay (ELISA) and immunochromatographic analysis, which are easy to perform and provide results within hours or even minutes [4][5][6][7]. However, immunogenic bacterial membrane proteins are required to ensure immunochemical test specificity.…”

Section: Introductionmentioning

confidence: 99%

Recombinant Salmonella enterica OmpX protein expression and its potential for serologically diagnosing Salmonella abortion in mares

Borovikov,

Ryskeldina,

Tursunov

et al. 2023

Vet World

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Background and Aim: Salmonella abortion in mares is caused by Salmonella enterica subspecies enterica serovar abortus equi infection and is characterized by premature (abortion) or non-viable fetus birth. Although all horses are susceptible to infection, the condition is more often clinically manifested in pregnant mares, with most abortions recorded in young females. In addition, nonspecific clinical disease signs and poorly sensitive and effective bacteriological diagnostic methods hinder rapid and reliable infection diagnoses. Immunochemical methods such as enzyme-linked immunosorbent assay (ELISA) and immunochromatography assays can facilitate effective and rapid diagnoses. However, they require highly specific and active antigens and antibodies. This study aimed to generate a recombinant S. enterica outer membrane protein X (OmpX) and evaluate its suitability for serological diagnosis of Salmonella abortion in mares. Materials and Methods: Outer membrane protein X from the S. enterica antigen was synthesized de novo and expressed in Escherichia coli using the pET28 vector. Transformed E. coli cells were cultured under different conditions to detect recombinant OmpX (rOmpX) expression, and rOmpX purification and refolding were both conducted using metal affinity chromatography. Refolded and purified rOmpX was characterized by western blotting, liquid chromatography with tandem mass spectrometry, and ELISA. Results: After optimized rOmpX expression, a 23 kDa molecular weight protein was identified. Amino acid sequence analysis using Mascot program suggested that these peptides were the OmpX protein from S. enterica. High specificity and diagnostic efficiency were recorded when rOmpX was used in ELISA against 89 serum samples from aborted and contact mares. Conclusion: Recombinant outer membrane protein, in comparison to the O antigen, demonstrated better diagnostic characteristics against sera from mares who aborted and contact horses. Keywords: diagnostics, outer membrane proteins, recombinant antigens, Salmonella abortion in mares, Salmonella enterica, specific antibodies.

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“…The impact on human health is significant as pathogenic bacterial strains are accountable for a diverse array of infectious diseases in humans. Notable instances comprise Staphylococcus aureus, which leads to skin and wound inflammation [8,9], Salmonella, which results in food poisoning [10,11], Escherichia coli, which causes infections of the bladder and gastrointestinal disorders [12][13][14][15], and Streptococcus pneumoniae, which causes respiratory infections and meningitis [16,17]. The severity of infections can vary, and inadequate treatment may result in life-threatening consequences.…”

mentioning

confidence: 99%

Electrochemical and optical-based systems for SARS-COV-2 and various pathogens assessment

Ahmed,

Ansari,

Siddiqui

et al. 2023

Adv. Nat. Sci: Nanosci. Nanotechnol.

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A critical step in the process for preventing and identifying emergencies relating to health, safety, and welfare is the testing and quick diagnosis of microbial pathogens. Due to the fast spread of waterborne and food borne infections in society and the high costs associated with them, pathogen identification has emerged as one of the most difficult parts of the water and food sectors. Since the turn of the century, pathogens have demonstrated enormous epidemiological and pandemic potential. The emergence and dissemination of a novel virus with pandemic potential endanger the livelihoods and well-being of individuals worldwide. The severe acute respiratory syndrome-coronavirus-2 (SARS-COV-2) coronavirus pandemic has propagated to almost every country on Earth and has had a considerable negative influence on economies and communities. Despite improvements in identification techniques for viral diseases, all nations must now execute biosensing in a speedy, sensitive, focused, and consistent manner in order to address pressing global issues. Hence, in this review, we have critically summarised the recent advancement of electrochemical as well as optical biosensors for the monitoring of SARS-COV-2 and various pathogens. Then, we began by providing a technical overview of cutting-edge strategies utilised to combat diseases and emergencies for it, including the utilisation of point-of-care technology (POCT), artificial intelligence (AI), and the internet of medical things (IoMT). This review article explores the integration of POC, IoMT, and AI technologies in the context of personal healthcare, focusing on their potential to expedite the diagnosis and treatment of medical conditions, ultimately leading to improved patient outcomes. Subsequently, the notion and execution of multiplex testing are presented to enhance the comprehension of detecting multiple analytes. Finally, conclusions and future directions have been presented.

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Development and Application of Real-Time PCR Assay for Detection of Salmonella Abortusequi

Abstract: Salmonella enterica subsp. enterica serovar Abortusequi is a major pathogen in horse and donkey herds, causing abortion in pregnant equids and resulting in enormous economic losses. A rapid and reliable method is urgently needed to detect S .

Cited by 9 publications

References 31 publications

Development and Application of an iELISA for the Detection of Antibody against Salmonella Abortusequi

Development and Application of an iELISA for the Detection of Antibody against Salmonella Abortusequi

Recombinant Salmonella enterica OmpX protein expression and its potential for serologically diagnosing Salmonella abortion in mares

Electrochemical and optical-based systems for SARS-COV-2 and various pathogens assessment

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