False‐negative results by polymerase chain reaction due to contamination by glove powder

Lomas, Juan García de; Sunzeri, Franklin J.; Busch, MP

doi:10.1046/j.1537-2995.1992.32192116439.x

Cited by 38 publications

(17 citation statements)

References 16 publications

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“…The ability to detect phage DNA was not obviously hindered by the presence of glove powder, as data for powdered and nonpowdered gloves among the gloves evaluated were comparable (Table 1), an important finding given the current prevalence of powdered gloves in use. There is at least one report that glove powder can inhibit PCR (4). In that study powder from a laboratory worker's gloves evidently entered PCR tubes during handling and subsequently inhibited DNA amplification.…”

Section: Discussionmentioning

confidence: 99%

PCR-Based Method for Detecting Viral Penetration of Medical Exam Gloves

2002

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The test approved by the U.S. Food and Drug Administration for assessment of the barrier quality of medical exam gloves includes visual inspection and a water leak test. Neither method tests directly the ability of gloves to prevent penetration by microorganisms. Methods that use microorganisms (viruses and bacteria) to test gloves have been developed but require classical culturing of the organism to detect it. We have developed a PCR assay for bacteriophage X174 that allows the rapid detection of penetration of gloves by this virus. The method is suitable for use with both latex and synthetic gloves. The presence of glove powder on either latex or synthetic gloves had no effect on the ability of the PCR assay to detect bacteriophage DNA. The assay is rapid, sensitive, and inexpensive; requires only small sample volumes; and can be automated.In response to a growing need to protect health care workers and patients from the possibility of transmission of human immunodeficiency virus through contact with body fluids, the Occupational Safety and Health Administration issued standard precautions that state that gloves must be worn when "touching blood, body fluids, secretions, excretions, and contaminated items" (17). Consequently, the rate of use of exam gloves in clinical practice rose dramatically (7). To ensure consistent quality in the manufacture of gloves, the U.S. Food and Drug Administration (FDA) introduced a two-part testing protocol in which gloves are first visually inspected and then subjected to a water leak test (5). These tests, while convenient and inexpensive, suffer from a number of drawbacks. Both the visual test for defects and the water leak test, in which an inspector looks for leaks in a glove filled with a liter of water, are subject to operator error. It has also been observed that microorganisms can penetrate gloves that do not show visible water leaks (2, 9). Studies that use microorganisms to gauge the propensities of gloves to permit pathogen penetration examine the question more directly than water leak studies. However, the microbial assays described rely on classical microbiological culture techniques to detect microbial penetration and therefore are more labor-intensive, time-consuming, and expensive than the FDA-approved tests. This report describes an assay for the detection of glove penetration by a small virus, bacteriophage X174. The assay uses PCR to detect the bacteriophage. PCR offers the advantages of being highly sensitive and specific for the agent being detected. With the recent and rapid increase in the availability of laboratory robotics for molecular biology, this PCR assay also has the potential to be automated and therefore made useful in the glove manufacturing setting. Cultures of E. coli were maintained on solid NB medium, made as described above by the addition of 15 g of agar per liter and 1 ml of CaCl 2 and the omission of Tween 80. For plaque analysis of bacteriophage X174, the top agar composition was identical to that of the plate agar, except that the co...

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Section: Discussionmentioning

confidence: 99%

PCR-Based Method for Detecting Viral Penetration of Medical Exam Gloves

2002

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“…Powder contamination from powdered gloves may give rise to sporadic false-negative PCR reactions (49 ).…”

Section: Document If Blood Taken From IV Linementioning

confidence: 99%

Preanalytical Variables Affecting the Integrity of Human Biospecimens in Biobanking

Ellervik

Vaught²

2015

Clinical Chemistry

139

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BACKGROUND Most errors in a clinical chemistry laboratory are due to preanalytical errors. Preanalytical variability of biospecimens can have significant effects on downstream analyses, and controlling such variables is therefore fundamental for the future use of biospecimens in personalized medicine for diagnostic or prognostic purposes. CONTENT The focus of this review is to examine the preanalytical variables that affect human biospecimen integrity in biobanking, with a special focus on blood, saliva, and urine. Cost efficiency is discussed in relation to these issues. SUMMARY The quality of a study will depend on the integrity of the biospecimens. Preanalytical preparations should be planned with consideration of the effect on downstream analyses. Currently such preanalytical variables are not routinely documented in the biospecimen research literature. Future studies using biobanked biospecimens should describe in detail the preanalytical handling of biospecimens and analyze and interpret the results with regard to the effects of these variables.

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“…Not all of the analysed cDNA samples showed unexpected transcripts. This could be due to unoptimized RT-PCR conditions without a nested strategy to enhance yield and specificity (Aittomaki et al, 1995;Roberts et al, 1993), a low RT-PCR cDNA input, and the low sensitivity of the ethidium bromide staining or other less frequent reasons for false-negative PCR results (de Lomas et al, 1992). Furthermore, genomic DNA contamination which can never be completely eliminated could compete for primers and dNTPs giving a negative result.…”

Section: Discussionmentioning

confidence: 99%

Transcription of thyroid autoantigens in non-expressing tissues

Aust

Crisp²,

Bösenberg³

et al. 2009

Exp Clin Endocrinol Diabetes

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Reverse transcriptase- polymerase chain reaction (RT-PCR) enhances the probability of detecting rare transcripts in complex mixtures of mRNA. Using thyroid autoantigens and the controversy about the role of the TSH-receptor (TSH-R) in thyroid-associated ophthalmopathy as an example, this study demonstrates the problems of interpreting RT-PCR results in typically non-expressing tissues resulting from the extremely high sensitivity of the method. Unexpected transcripts for thyroperoxidase, thyroglobulin, TSH-R (exon 1-4, 354 bp), FSH-receptor, or insulin fragments were demonstrated in a number of thyroid or orbit-derived as well as unrelated tissues or cell types. Unexpected transcripts were most prevalent in fibroblasts, irrespective of the tissue of origin and most likely caused by ectopic transcription. To establish a physiological significance of rare transcripts such as the TSH-R in orbital tissues, demonstration of the protein in addition to the positive RT-PCR results is needed.

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False‐negative results by polymerase chain reaction due to contamination by glove powder

Cited by 38 publications

References 16 publications

PCR-Based Method for Detecting Viral Penetration of Medical Exam Gloves

PCR-Based Method for Detecting Viral Penetration of Medical Exam Gloves

Preanalytical Variables Affecting the Integrity of Human Biospecimens in Biobanking

Transcription of thyroid autoantigens in non-expressing tissues

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