Inhibitors of RT-PCR in serum

Konet, D.S; Mezêncio, J. M. S.; Babcock, Gwen D.; Brown, F.

doi:10.1016/s0166-0934(99)00131-7

Cited by 18 publications

(11 citation statements)

References 4 publications

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“…Clearly, the RNA extraction method is most important. Many PCR inhibitors are presumed to affect directly the RNA or the enzymes of the reaction [20,25]. Several inhibitors have been found in faeces: polysaccharides or chlorophyll originating from vegetables, bile salts, urea, glycolipids and heparin [26][27][28].…”

Section: Discussionmentioning

confidence: 99%

Performance of a commercial assay for detecting and quantifying HEV RNA in faeces

Abravanel

Lacipière

Lhomme

et al. 2018

Journal of Clinical Virology

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A B S T R A C TBackground: Detecting hepatitis E virus (HEV) RNA in faeces is useful for diagnosing and monitoring HEV infections, particularly in immunocompromised patients requiring ribavirin therapy. Objectives: This study evaluated the performance of the Altona RealStar HEV RNA kit for detecting and quantifying HEV in faeces. Study design: RNA was extracted from 94 stool samples by two methods: QIAamp Viral RNA Mini kit and MagNA Pure 96 automate. The Altona results were compared to a reference laboratory-developed accredited ISO15189 RT-PCR assay. Results: The Altona and reference assays detect HEV RNA in 77/93 (82.8%) and 83/93 (89.2%) of the QIAamp extracted samples, respectively, after exclusion of invalid result; they detected HEV RNA in 67/92 (72.8%) and 66/92 (71.7%) of the MagNA Pure extracted samples, respectively, which emphasizes the importance of the RNA extraction method. The HEV RNA concentrations obtained with Altona RT-PCR and the reference RT-PCR were well correlated whatever the extraction method, and Bland Altman analyses indicated that the Altona values were higher than the reference assay values. The Altona values for QIAamp-extracted and MagNA Pure-extracted HEV RNA were very similar. Conclusions: The Altona RealStar assay is suitable for quantifying HEV RNA in the faeces and monitoring HEV RNA shedding during ribavirin therapy. Extraction is critical for detecting faecal HEV with high performance RT-PCR assays.

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Section: Discussionmentioning

confidence: 99%

Performance of a commercial assay for detecting and quantifying HEV RNA in faeces

Abravanel

Lacipière

Lhomme

et al. 2018

Journal of Clinical Virology

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“…Many of these inhibitors are presumed to affect directly the RNA and not the enzymes of the reaction (Konet et al . ). In addition, hormones or antiviral substances like acyclovir can also affect amplification (Yedidag et al .…”

Section: Pcr Inhibitors In Clinical Samplesmentioning

confidence: 97%

PCR inhibitors - occurrence, properties and removal

et al. 2012

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Summary The polymerase chain reaction (PCR) is increasingly used as the standard method for detection and characterization of microorganisms and genetic markers in a variety of sample types. However, the method is prone to inhibiting substances, which may be present in the analysed sample and which may affect the sensitivity of the assay or even lead to false‐negative results. The PCR inhibitors represent a diverse group of substances with different properties and mechanisms of action. Some of them are predominantly found in specific types of samples thus necessitating matrix‐specific protocols for preparation of nucleic acids before PCR. A variety of protocols have been developed to remove the PCR inhibitors. This review focuses on the general properties of PCR inhibitors and their occurrence in specific matrices. Strategies for their removal from the sample and for quality control by assessing their influence on the individual PCR test are presented and discussed.

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“…PCR inhibitors found in blood, serum or plasma samples includes substances like IgG, haemoglobin and lactoferrin (Al-Soud et al, 2000;Al-Soud et al, 2001). Most of these inhibitors are presumed to affect the RNA directly and not the enzymes of the reaction (Konet et al, 2000).…”

Section: Diagnosis Of Bovine Tuberculosis In Live Animals Ismentioning

confidence: 99%

Detection and Identification of Mycobacterium tuberculosis and Mycobacterium bovis from Blood and Milk of Bovines

Gill¹

2020

JAR

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2020). Detection and identification of Mycobacterium tuberculosis and Mycobacterium bovis from blood and milk of bovines. J. Anim. Res., 10(1): 59-65. ABSTRACTBovine tuberculosis, a chronic disease of animals is caused by species of Mycobacterium tuberculosis complex (MTC) and it remains a potential threat to animals as well as humans. Differentiation of the species of MTC is required for epidemiological and diagnostic purpose. The present study evaluated the presence of different species of MTC in bovines using gyrB-restriction fragment length polymorphism analysis. In this study, blood and milk samples from 50 milch animals which were positive reactors of comparative intradermal tuberculin test were collected. Screening of MTC was done by IS6110-PCR using primers INS1/INS2 specific for MTC. The positive samples were further identified using gyrB-Restriction fragment length polymorphism analysis. Out of 50 positive reactors to CITT, only 4 (8%) animal were positive for MTC by IS6110-PCR. And gyrB-RFLP analysis using RsaI and SacII showed two positive for M. bovis and two animals for M. tuberculosis. Thus, gyrB-RFLP could be used as an additional tool in differential diagnosis of mycobacterial diseases thereby able to differentiate species of MTC.

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Inhibitors of RT-PCR in serum

Cited by 18 publications

References 4 publications

Performance of a commercial assay for detecting and quantifying HEV RNA in faeces

Performance of a commercial assay for detecting and quantifying HEV RNA in faeces

PCR inhibitors - occurrence, properties and removal

Detection and Identification of Mycobacterium tuberculosis and Mycobacterium bovis from Blood and Milk of Bovines

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