False-Positive Results of Plasma PCR for Cytomegalovirus DNA due to Delayed Sample Preparation

Schäfer, Peter; Tenschert, W; Schröter, Matthias; Gutensohn, K.; Laufs, Rainer

doi:10.1128/jcm.38.9.3249-3253.2000

Cited by 30 publications

(5 citation statements)

References 21 publications

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“…The relatively large number of CMV by PCR in blood of immune-competent children were most likely not all acute cases even though primary infection with CMV occurs during the first year of life 15 . A certain proportion of the PCR-positives might be explained by delayed sample preparation and consequently, the release of leukocyte-derived CMV DNA 16 . EBV is rather expected to be seen in young adults than in young children 17 .…”

Section: Discussionmentioning

confidence: 99%

Causes of fever in Gabonese children: a cross-sectional hospital-based study

Fernandes

Held

Dorn

et al. 2020

Sci Rep

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The causes of infections in pediatric populations differ between age groups and settings, particularly in the tropics. Such differences in epidemiology may lead to misdiagnosis and ineffective empirical treatment. Here, we investigated the current spectrum of pathogens causing febrile diseases leading to pediatric hospitalization in Lambaréné, Gabon. From August 2015 to March 2016, we conducted a prospective, cross-sectional, hospital-based study in a provincial hospital. Patients were children ≤ 15 years with fever ≥ 38 °C and required hospitalization. A total of 600 febrile patients were enrolled. Malaria was the main diagnosis found in 52% (311/600) patients. Blood cultures revealed septicemia in 3% (17/593), among them four cases of typhoid fever. The other causes of fever were heterogeneously distributed between both bacteria and viruses. Severe infections identified by Lambaréné Organ Dysfunction Score (LODS) were also most often caused by malaria, but children with danger signs did not have more coinfections than others. In 6% (35/600) of patients, no pathogen was isolated. In Gabon, malaria is still the major cause of fever in children, followed by a bacterial and viral disease. Guidelines for both diagnosis and management should be tailored to the spectrum of pathogens and resources available locally.Causes of fever in African pediatric populations are more diverse than previously thought. A landmark study conducted in Tanzania showed that due to a change in epidemiology, a broad spectrum of pathogens replaced P. falciparum malaria as the most common cause of disease in children in this area 1 . However, a few years later, P. falciparum malaria, is still seen to be the main cause of febrile illnesses in Ghana, West Africa 2 . When unaware, these differences in epidemiology might lead to misdiagnosis as well as inefficient treatment by the medical personnel. The process of medical diagnosis includes the joint interpretation of symptoms, clinical signs and laboratory findings. Careful selection and prioritization of a diagnostic setup are informed by a priori knowledge of the seasonal, local and worldwide frequency and distribution of a given disease 3,4 .Our study describes the distribution of infections, co-infections, and co-morbidities in children hospitalized for febrile illnesses at the Albert Schweitzer Hospital (HAS) in Lambaréné, Gabon, as an example for a hospital in a semiurban Central African region. In addition, we present the current spectrum of pathogens causing severe disease identified by Lambaréné Organ Dysfunction Score (LODS) in these children. ResultsStudy patients. A total of 600 febrile patients ≤ 15 years were enrolled in our study. Of these, 280 (47%) were females; 69% (415/600) patients were < 5 years, and median (IQR) age was 29 [12-68] months (Table 1). Seven percent (40/549, NA = 51) had at least one known chronic medical condition prior to admission, among the main ones: 4% (23/600) patients had homozygous sickle cell disease; 1% (6/600) were HIV positive. Vaccination coverage...

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Section: Discussionmentioning

confidence: 99%

Causes of fever in Gabonese children: a cross-sectional hospital-based study

Fernandes

Held

Dorn

et al. 2020

Sci Rep

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“…The detection of viral infections plays an important role in clinical diagnosis and monitoring of drug therapy. Herpes virus diagnostics for example can be applied in the fields of transplant patient monitoring, encephalitis, neurological syndromes, opportunistic infections in immune-suppressed patients, or screenings of blood donations. − The molecular diagnosis of these viruses is commonly based on PCR due to the low abundance of viral DNA in biological samples. A multiplexed PCR format enabled the amplifying of multiple target sequences simultaneously in one reaction. ,, The analysis of the resulting amplicon mixtures is based on hybridization of target DNA employing an enzyme label as redox molecule producing molecular species. , The application of these enzyme-linked immunosorbent assay (ELISA) formats to fully electrical multiposition arrays is shown.…”

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confidence: 99%

“…Herpes virus diagnostics for example can be applied in the fields of transplant patient monitoring, encephalitis, neurological syndromes, opportunistic infections in immune-suppressed patients, or screenings of blood donations. [23][24][25][26] The molecular diagnosis of these viruses is commonly based on PCR due to the low abundance of viral DNA in biological samples. A multiplexed PCR format enabled the amplifying of multiple target sequences simultaneously in one reaction.…”

mentioning

confidence: 99%

Electrical Detection of Viral DNA Using Ultramicroelectrode Arrays

et al. 2003

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A fully electrical array for voltammetric detection of redox molecules produced by enzyme-labeled affinity binding complexes is shown. The electronic detection is based on ultramicroelectrode arrays manufactured in silicon technology. The 200-microm circular array positions have 800-nm-wide interdigitated gold ultramicroelectrodes embedded in silicon dioxide. Immobilization of oligonucleotide capture probes onto the gold electrodes surfaces is accomplished via thiol-gold self-assembling. Spatial separation of probes at different array positions is controlled by polymeric rings around each array position. The affinity bound complexes are labeled with alkaline phosphatase, which converts the electrochemically inactive substrate 4-aminophenyl phosphate into the active 4-hydroxyaniline (HA). The nanoscaled electrodes are used to perform a sensitive detection of enzyme activity by signal enhancing redox recycling of HA resulting in local and position-specific current signals. Multiplexing and serial readout is realized using a CMOS ASIC module and a computer-controlled multichannel potentiostat. The principle of the silicon-based electrical biochip array is shown for different experimental setups and for the detection of virus DNA in real unpurified multiplex PCR samples. The fast and quantitative electronic multicomponent analysis for all kinds of affinity assays is robust and particle tolerant.

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“…Alternately, the high abundance of HCMV leukocyte DNAemia in actively HCMV‐infected patients has led to the hypothesis that plasma DNAemia may be, at least in part, a result of peripheral blood lysis. This hypothesis would imply that for patients with HCMV leukocyte DNAemia, long‐term storage of whole blood samples before separation may cause the release of viral DNA into the plasma fraction, thus distorting PCR results [Schäfer et al, ].…”

Section: Discussionmentioning

confidence: 99%

Human herpesvirus infections of the central nervous system: Laboratory diagnosis based on DNA detection by nested PCR in plasma and cerebrospinal fluid samples

Rimério

Oliveira

Bonatelli

et al. 2015

Journal of Medical Virology

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Infections of the central nervous systems (CNS) present a diagnostic problem for which an accurate laboratory diagnosis is essential. Invasive practices, such as cerebral biopsy, have been replaced by obtaining a polymerase chain reaction (PCR) diagnosis using cerebral spinal fluid (CSF) as a reference method. Tests on DNA extracted from plasma are noninvasive, thus avoiding all of the collateral effects and patient risks associated with CSF collection. This study aimed to determine whether plasma can replace CSF in nested PCR analysis for the detection of CNS human herpesvirus (HHV) diseases by analysing the proportion of patients whose CSF nested PCR results were positive for CNS HHV who also had the same organism identified by plasma nested PCR. In this study, CSF DNA was used as the "gold standard," and nested PCR was performed on both types of samples. Fifty-two patients with symptoms of nervous system infection were submitted to CSF and blood collection. For the eight HHV, one positive DNA result-in plasma and/or CSF nested PCR-was considered an active HHV infection, whereas the occurrence of two or more HHVs in the same sample was considered a coinfection. HHV infections were positively detected in 27/52 (51.9%) of the CSF and in 32/52 (61.5%) of the plasma, difference not significant, thus nested PCR can be performed on plasma instead of CSF. In conclusion, this findings suggest that plasma as a useful material for the diagnosis of cases where there is any difficulty to perform a CSF puncture.

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False-Positive Results of Plasma PCR for Cytomegalovirus DNA due to Delayed Sample Preparation

Cited by 30 publications

References 21 publications

Causes of fever in Gabonese children: a cross-sectional hospital-based study

Causes of fever in Gabonese children: a cross-sectional hospital-based study

Electrical Detection of Viral DNA Using Ultramicroelectrode Arrays

Human herpesvirus infections of the central nervous system: Laboratory diagnosis based on DNA detection by nested PCR in plasma and cerebrospinal fluid samples

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