Development of a routine laboratory direct detection system of staphylococcal enterotoxin genes

Nakayama, Akifumi; Okayama, Akira; Hashida, Misao; Yamamoto, Yasuzumi; Takebe, Hisakatsu; Ohnaka, Takashi; Tanaka, Tomoyuki; Imai, Shunsuke

doi:10.1099/jmm.0.46027-0

Cited by 16 publications

(19 citation statements)

References 26 publications

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“…The past reports from Germany suggest that Ct for fem B gene was between 21.8-33.7 and 22.5-33.5 for Se a gene [29]. Nakayama et al [9] detected eight Se-encoding genes in the S. aureus of milk sample, and the Ct value detection limit of Se encoding gene was between 39.44 and 37.92.…”

Section: Resultsmentioning

confidence: 99%

“…However, RT-PCR method of direct detection of Se from the sample is more sensitive and rapid than that of RPLA method. Furthermore, the RT-PCR method was also proved to be a successfully applicable technique for direct detection of Se from the clinical samples that contain high levels of PCR inhibitors [9]. Rajkovic also compared iqPCR with inhouse ELISA technique for detecting Se b toxin genes in both pure culture and contaminated food sample and found that iqPCR was approximately 1,000 times more sensitive than the in-house ELISA method [25].…”

Section: Resultsmentioning

confidence: 99%

“…However, Nakayama et al a rapid and direct detection of SE-encoding genes from food sample using a real-time polymerase chain reaction (RT-PCR) for routine laboratory analysis [9]. In this work, we have used RT-PCR to evaluate the existence SE-encoding genes in S. aureus isolated from subclinical mastitis milk sample; further, Se genes were also directly detected from the subclinical mastitis milk sample.…”

Section: Introductionmentioning

confidence: 99%

“…DNA extraction from the milk sample was carried out as per the method developed by Nakayama et al [9]. In this procedure, 1:1 volume milk sample (100 ml) and 0.2 M sodium hydroxide was taken and stored at 37°C for 20 min.…”

Section: Milk Sample Preparation For Qrt-pcrmentioning

confidence: 99%

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Rapid and sensitive method for detection of Staphylococcus aureus enterotoxin genes in milk sample

2018

J App Biol Biotech

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Staphylococcus aureus is a common microflora present on skin and mucous membrane of cattle. Despite its amiable nature, S. aureus can turn shrewd and get to be distinctly pathogenic. Staphylococcus present in the milk may produce numerous potential destructive variables, like toxins. Consequently, consumption of S. aureus contaminant milk may result in Staphylococcal food poisoning (SFP). Therefore, the present study was carried out to evaluate the presence of Se-encoding genes in S. aureus and also validate the detection of toxin genes directly from the milk sample contaminated with S. aureus using real-time polymerase chain reaction (RT-PCR). The RT-PCR results were reported as Ct values, which confirmed the presence of enterotoxin genes in the S. aureus isolates and milk samples. Application of RT-PCR for the detection of Se genes in cultures of S. aureus and also from milk samples revealed the probable onset of SFP. Therefore, the study suggests that the analysis of milk for the presence of any toxin genes should be strictly performed before reaching the consumer.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Milk Sample Preparation For Qrt-pcrmentioning

confidence: 99%

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Rapid and sensitive method for detection of Staphylococcus aureus enterotoxin genes in milk sample

2018

J App Biol Biotech

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“…Milchpulver entwickelt. Nakayama et al (2006) konnten sea -sei in künstlich kontaminierter Milch mit Nachweisgrenzen von 10 2 -10 4 KbE/ml nachweisen. Ikeda et al (2005b) untersuchten in Zusammenhang mit einem Ausbruch durch SEA-bildende S. aureus Milchpulver quantitativ auf sea, um damit indirekt auf die Belastung mit S. aureus zu schließen.…”

Section: Polymerasekettenreaktionunclassified

Staphylokokken-Enterotoxine: Bildung, Eigenschaften und Nachweis

2007

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Zusammenfassung: Staphylococcus aureus kann sich unter geeigneten Bedingungen in Lebensmitteln vermehren und dabei Enterotoxine bilden, die beim Konsumenten Erbrechen und Durchfall auslösen. Im Gegensatz zu dem Erreger selbst sind diese Toxine sehr widerstandsfähig insbesondere gegenüber hohen Temperaturen und können durch eine küchenmäßige Bearbeitung des sie enthaltenden Lebensmittels im allgemeinen nicht mehr inaktiviert werden. Da demnach die Anwesenheit der Staphylokokken zur Auslösung der Erkrankung nicht mehr erforderlich ist, handelt es sich bei dieser um eine typische Lebensmittelintoxikation. Die Abschätzung einer potentiellen Gesundheitsgefährdung über den Erregernachweis ist daher nur bei rohen Lebensmitteln sinnvoll. Bei Produkten, die im Verlauf ihrer Herstellung einem für die Keime abträglichen Verfahren, z. B. einer Wärmebehandlung, unterzogen wurden, kann der Thermonukleasetest (indirekter Nachweis einer Kontamination mit hoher Anzahl an Staphylokokken, unabhängig davon, ob die Erreger zum Zeitpunkt der Untersuchung noch präsent sind) oder der direkte Enterotoxinnachweis durchgeführt werden. Der Nachweis der Toxingene mit molekularbiologischen Verfahren hat in diesem Zusammenhang nur unterstützende Funktion, da im Fall eines positiven Ergebnisses aus diesem nicht geschlossen werden kann, daß die Gene auch exprimiert wurden, also Toxine im Lebensmittel vorhanden sind. In der vorliegenden Übersichtsarbeit werden der Erreger und seine Enterotoxine sowie die verschiedenen Nachweisverfahren unter besonderer Berücksichtigung der molekularbiologischen Methoden besprochen.Abstract: Under appropriate conditions, Staphylococcus aureus can grow in food and produce enterotoxins which cause vomiting and diarrhoea when ingested. In contrast to the staphylococci these toxins are resistant especially to high temperatures and are not inactivated by usual kitchen practice. As the presence of staphylococci is not essential to cause illness, this condition is a typical food poisoning. The estimation of a potential health risk by targeting the organism is therefore only appropriate to raw food. For products which underwent a procedure detrimental to the bacteria, e. g. heat treatment, testing for thermonuclease (indirect detection of a contamination with high numbers of staphylococci, irrespective of the presence of living bacteria at the time of testing) or the detection of enterotoxins can be applied. In this context, the detection of enterotoxin genes using molecular biological tests provides only supplemental information, as positive results are not evidentiary for gene expression and thus for the presence of toxins in the food. In this review, the causative organism and its toxins as well as methods of detection are discussed with special emphasis on molecular biological methods.

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Epidemiological survey of bovine gammaherpesvirus 4 (BoHV-4) infection in cattle and buffalo from Pakistan

et al. 2022

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Development of a routine laboratory direct detection system of staphylococcal enterotoxin genes

Cited by 16 publications

References 26 publications

Rapid and sensitive method for detection of Staphylococcus aureus enterotoxin genes in milk sample

Rapid and sensitive method for detection of Staphylococcus aureus enterotoxin genes in milk sample

Staphylokokken-Enterotoxine: Bildung, Eigenschaften und Nachweis

Epidemiological survey of bovine gammaherpesvirus 4 (BoHV-4) infection in cattle and buffalo from Pakistan

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