A Multiplex PCR Assay Mediated by Universal Primer for the Diagnosis of Human Meningitis Caused by Six Common Bacteria

Lu, Sangwei; Hu, Yingxi; Shen, Songdong; Xi, Bangsheng; Wang, X. N.; Sun, Wanping

doi:10.1134/s1022795418040075

Cited by 4 publications

(2 citation statements)

References 34 publications

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“…Our universal multiplex protocol is rapid (detecting up to four genes in a single PCR run), easy to employ, and cost-effective because no specialized laboratory equipment is required. Our results are consistent with the findings of Duan et al [ 54 ], who reported a high-specificity universal multiplex PCR assay capable of detecting multiple pathogens associated with human bacterial meningitis. We believe that agarose-gel-based multiplex PCR is one of the best approaches for genotyping a large number of parents (varieties), progeny, and/or weedy rice populations in molecular laboratories with limited resources.…”

Section: Discussionsupporting

confidence: 93%

Novel PCR-Based Multiplex Assays for Detecting Major Quality and Biotic Stress in Commercial and Weedy Rice

Hanafiah

Cheng

Lim

et al. 2022

Life

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While previous research has demonstrated that multiplex polymerase chain reaction (PCR) can be a cost-effective approach to detect various genes in crops, the availability of multiplex assays to simultaneously screen both grain quality and biotic stress resistance traits in rice (Oryza sativa) is limited. In this work, we report six novel multiplex assays that use a universal protocol to detect major rice grain quality (amylose content and fragrance) and biotic stress (blast, sheath blight, and bacterial leaf blight) traits with amplified products consisting of up to four primer pairs that can be analyzed using a standard agarose-based gel electrophoresis system. Recent studies have suggested that weedy rice has novel sources of disease resistance. However, an intensive screening of weedy biotypes has not been reported in Malaysia. Accordingly, we employed one of the developed multiplex assays to screen reported genes or quantitative trait loci (QTLs) associated with blast, sheath blight, and bacterial leaf blight diseases in 100 weedy rice biotypes collected from five local fields, with phenotyping performed to validate the genotyping results. In conclusion, our universal multiplex protocol is effective for the large-scale genotyping of rice genetic resources, and it can be employed in routine molecular laboratories with limited resources.

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Section: Discussionsupporting

confidence: 93%

Novel PCR-Based Multiplex Assays for Detecting Major Quality and Biotic Stress in Commercial and Weedy Rice

Hanafiah

Cheng

Lim

et al. 2022

Life

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“…In the early 10 cycles, the low concentration of chimeric primers functioned to yield PCR products with common primer tags at a relatively higher annealing temperature first (66 °C). The PCR amplification products of the first 10 cycles containing the common primer sequence, which continue to be amplified 35 cycles by a high concentration of the common primer at a lower annealing temperature (53 °C; Duan et al., ). The CP‐MPCR was performed in a total volume of 25 µL containing 12.5 µL Taq Master Mix, 2 µL of chimeric primers mixture, 40 µM common primers, and 1 µL (20 ng) DNA template.…”

Section: Methodsmentioning

confidence: 99%

A multiplex PCR assay with a common primer for the detection of eleven foodborne pathogens

Tao

Liu

Ding

et al. 2020

Journal of Food Science

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Salmonella enterica, Listeria monocytogenes, Shigella flexneri, Escherichia coli O157:H7, Vibrio parahaemolyticus, Staphylococcus aureus, Vibrio cholerae, Clostridium botulinum type A, Bacillus cereus, Clostridium perfringens Alpha toxin, and Yersinia enterocolitica are 11 common foodborne pathogens. Traditional bacterial culture methods for detecting pathogens are time‐consuming and labor‐intensive. Multiplex PCR technology, which can detect multiple targets in a single tube, has been increasingly applied to microbial detection due to its high specificity, sensitivity, and fast response. This paper is to establish a multiplex PCR technology mediated by a common primer for the detection of these 11 common foodborne pathogens in order to achieve the goal of nondirectional screening for these 11 common foodborne pathogens. The specificity of the established CP‐MPCR detection system was first verified by 100 clinical isolates. The sensitivity of the CP‐MPCR detection system was then detected by using cultured bacteria preparations and has been confirmed with a high sensitivity of 103 to 104 CFU/mL, among them, the sensitivity of the CP‐MPCR for Vibrio cholerae and S. flexneri can even achieve 102 CFU/mL. Sixty anal swab samples collected from Suzhou CDC and 16 enrichment cultured solutions of food samples collected from the Suzhou Food Inspection and Testing Center were tested using the CP‐MPCR system. A total of 32 positive results were detected. Practical Application Food poisoning incidents occur frequently around the world, mainly because of the contamination of food by pathogenic bacteria and serious harm to human health. The method provided in this study can detect 11 foodborne pathogens in food, which can effectively prevent the spread of pathogenic microorganisms. At the same time, for the food poisoning incident that has already occurred, this method can be used for diagnosis to find out the cause.

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Universal primer multiplex PCR assay for detection and genotyping of porcine astroviruses

Zhang,

Wen,

Liu

et al. 2023

Journal of Virological Methods

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A Multiplex PCR Assay Mediated by Universal Primer for the Diagnosis of Human Meningitis Caused by Six Common Bacteria

Cited by 4 publications

References 34 publications

Novel PCR-Based Multiplex Assays for Detecting Major Quality and Biotic Stress in Commercial and Weedy Rice

Novel PCR-Based Multiplex Assays for Detecting Major Quality and Biotic Stress in Commercial and Weedy Rice

A multiplex PCR assay with a common primer for the detection of eleven foodborne pathogens

Universal primer multiplex PCR assay for detection and genotyping of porcine astroviruses

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