Epistemic_Statements  

CORD-19:6b4b32c9c9f84ee5d0a74196f5d9378a955adac1 JSONTXT 11 Projects

Annnotations TAB TSV DIC JSON TextAE

Id Subject Object Predicate Lexical cue
T1 994-1097 Epistemic_statement denotes An internal control ensures detection of efficient nucleic acid extraction and possible PCR inhibition.
T2 2331-2561 Epistemic_statement denotes Apart from the 6 C-terminal codons, the E1 gene is largely conserved among alphaviruses (Powers et al., 2001) , although there is sufficient diversity to allow selection of primers and probes that are specific for the EEEV genome.
T3 3134-3277 Epistemic_statement denotes Reports of case fatalities range from 5% to 20% (Tsai and Mitchell, 1989) , although the numbers are higher still among the elderly population.
T4 3780-4137 Epistemic_statement denotes However, serologic tests have certain limitations; cross-reactivity of flaviviral antibodies is problematic in immunoglobulin M capture enzyme-linked immunosorbent assays, and plaque reduction neutralization is time consuming, is cumbersome, and requires biosafety level 3 (BSL-3) containment (Calisher, 1994; Martin et al., 2004; Russell and Dwyer, 2000) .
T5 4138-4344 Epistemic_statement denotes The traditional method of virus isolation also has the latter limitations, and in addition, because the period of viremia is generally short, the likelihood of obtaining an isolate is low (Calisher, 1994) .
T6 8205-8511 Epistemic_statement denotes The advantage of using RNA transcript rather than genomic RNA as the positive control is that the RNA transcript, unlike the genomic EEEV RNA, can be handled in a BSL-2 laboratory that is not SA certified by non-SA-approved personnel; diagnostic testing for EEEV is generally performed in BSL-2 facilities.
T7 13224-13434 Epistemic_statement denotes Because of the SA status of EEEV, a positive control for the assay was constructed that could be used in a BSL-2 laboratory by non-SA-approved personnel, thereby facilitating the assay's diagnostic application.
T8 15418-15529 Epistemic_statement denotes The assay can detect a range between 5 and 5 × 10 6 gc of EEEV and a range between 10 and 3 × 10 6 gc for SLEV.
T9 15654-15927 Epistemic_statement denotes The assay range and sensitivity of the duplex assay, when compared with the SLE and EEE assays performed separately as singleplex assays, gave similar results, indicating that there is no negative interference between the primer probe sets of the 2 assays (data not shown).
T10 17291-17514 Epistemic_statement denotes We then performed the singleplex real-time RT-PCR assay for the detection of EEEV on various RNA dilutions to determine whether nucleic acid from the 12 different strains would be amplified by the real-time RT-PCR reaction.
T11 19218-19391 Epistemic_statement denotes For each of the blinded samples, the high, medium, and low gc samples could all be detected indicating minimal loss in sensitivity after nucleic acid extraction ( Table 4 ).
T12 19392-19581 Epistemic_statement denotes All of the negative extraction controls had Ct values of N45, indicating that false positives were not detected and that no crosscontamination had occurred during extraction of the samples.
T13 20073-20227 Epistemic_statement denotes In general, if a clinical specimen showed a higher than acceptable GFP Ct value, the recommendation would be to repeat the extraction and realtime RT-PCR.
T14 20926-21033 Epistemic_statement denotes Human infections by SLEV and EEEV are relatively rare, although EEEV infection of equines occurs regularly.
T15 21034-21158 Epistemic_statement denotes Nevertheless, severe morbidity and mortality associated with these 2 arboviruses make them important public health concerns.
T16 22393-22771 Epistemic_statement denotes SLEV, in contrast, is found throughout the 48 contiguous states, but periodic epidemics have only been documented in the Midwest and the Southeastern United States (including Table 4 Average Ct values obtained when the EEEV, SLEV, and GFP real-time RT-PCR assays were performed on extracts of CSF specimens spiked with SLEV culture and EEEV transcript NTC = no template control.
T17 23183-23431 Epistemic_statement denotes A molecular assay that detects both SLEV and EEEV, thus, has considerable application for patients suspected of having arboviral encephalitis who reside in these areas, as well as travelers who could have come into contact with infected mosquitoes.
T18 23670-23763 Epistemic_statement denotes The assay is intended for patient CSF samples but can easily be used for vector surveillance.
T19 24470-24725 Epistemic_statement denotes Although the 2 assays were similar in sensitivity and specificity, the previously published assay did not detect all SLEV strains; 4 of the 69 SLEV strains in Table 2 (CorAn 9275, CorAn 9124, GML 903797, and GML 903369) were not detected (data not shown).
T20 25466-25641 Epistemic_statement denotes Nevertheless, detection of both viral genomes occurred just as efficiently as if there was no competitor virus present (Table 4 ), indicating that neither assay was inhibited.
T21 25759-25991 Epistemic_statement denotes It should be noted that, in our experience, after having tested well more than 1000 CSF specimens, CSF specimens rarely (b1%) exhibit significant PCR inhibition, whereas a larger number of plasma and serum specimens show inhibition.
T22 25992-26283 Epistemic_statement denotes We would, therefore, recommend that if plasma or serum specimens are tested, the nucleic acid extracts from the specimens are not diluted before performing the GFP real-time RT-PCR assay to detect inhibition more accurately because the predilution may overcome some of the inhibitory effect.