Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.49 (reverse transcriptase)
 31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Laboratory diagnosis of imported, vector-borne virus diseases during a 22-month-period in Munich, Germany, is summarized. IN 13/317 Germans returning from the Mediterranean with suspected sandfly fever, acute sandfly fever, serotype Toscana, was confirmed serologically: 84.6% of the infections were acquired in Italy. Of 249 German tourists with febrile disease returning from the tropics, acute infection with dengue virus was diagnosed serologically in 26 (10.4%): most infections were acquired in Thailand (57.7%). In a seroepidemiological study of 670 German aid workers who had spent two years in the tropics, 49 (7.3%) were positive for antibodies to dengue, 9 (1.3%) to chikungunya, and 1 (0.1%) to Sindbis virus. Of 17 Middle Eastern patients with suspected viral haemorrhagic fever, genomic Crimean-Congo haemorrhagic fever virus RNA was amplified in 4 (23.5%) by semi-nested reverse transcriptase polymerase chain reaction, and confirmed by molecular characterization of nucleic acid. With the increase in travel to and from endemic areas, imported vector-borne virus infections are increasingly important in Germany.
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PMID:Travel-related vector-borne virus infections in Germany. 880 Aug 6

Viral hemorrhagic fevers (VHFs) are acute infections with high case fatality rates. Important VHF agents are Ebola and Marburg viruses (MBGV/EBOV), Lassa virus (LASV), Crimean-Congo hemorrhagic fever virus (CCHFV), Rift Valley fever virus (RVFV), dengue virus (DENV), and yellow fever virus (YFV). VHFs are clinically difficult to diagnose and to distinguish; a rapid and reliable laboratory diagnosis is required in suspected cases. We have established six one-step, real-time reverse transcription-PCR assays for these pathogens based on the Superscript reverse transcriptase-Platinum Taq polymerase enzyme mixture. Novel primers and/or 5'-nuclease detection probes were designed for RVFV, DENV, YFV, and CCHFV by using the latest DNA database entries. PCR products were detected in real time on a LightCycler instrument by using 5'-nuclease technology (RVFV, DENV, and YFV) or SybrGreen dye intercalation (MBGV/EBOV, LASV, and CCHFV). The inhibitory effect of SybrGreen on reverse transcription was overcome by initial immobilization of the dye in the reaction capillaries. Universal cycling conditions for SybrGreen and 5'-nuclease probe detection were established. Thus, up to three assays could be performed in parallel, facilitating rapid testing for several pathogens. All assays were thoroughly optimized and validated in terms of analytical sensitivity by using in vitro-transcribed RNA. The >or=95% detection limits as determined by probit regression analysis ranged from 1,545 to 2,835 viral genome equivalents/ml of serum (8.6 to 16 RNA copies per assay). The suitability of the assays was exemplified by detection and quantification of viral RNA in serum samples of VHF patients.
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PMID:Rapid detection and quantification of RNA of Ebola and Marburg viruses, Lassa virus, Crimean-Congo hemorrhagic fever virus, Rift Valley fever virus, dengue virus, and yellow fever virus by real-time reverse transcription-PCR. 1208 42

We developed four assays for specifically identifying Dobrava (DOB), Hantaan (HTN), Puumala (PUU), and Seoul (SEO) viruses. The assays are based on the real-time one-step reverse transcriptase polymerase chain reaction (RT-PCR) with the small segment used as the target sequence. The detection limits of DOB, HTN, PUU, and SEO assays were 25, 25, 25, and 12.5 plaque-forming units, respectively. The assays were evaluated in blinded experiments, each with 100 samples that contained Andes, Black Creek Canal, Crimean-Congo hemorrhagic fever, Rift Valley fever and Sin Nombre viruses in addition to DOB, HTN, PUU and SEO viruses. The sensitivity levels of the DOB, HTN, PUU, and SEO assays were 98%, 96%, 92% and 94%, respectively. The specificity of DOB, HTN and SEO assays was 100% and the specificity of the PUU assay was 98%. Because of the high levels of sensitivity, specificity, and reproducibility, we believe that these assays can be useful for diagnosing and differentiating these four Old-World hantaviruses.
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PMID:Identification of Dobrava, Hantaan, Seoul, and Puumala viruses by one-step real-time RT-PCR. 1566 46

Crimean-Congo hemorrhagic fever (CCHF) is endemic in certain rural areas of Pakistan. Since the discovery of CCHF virus (CCHFV) in the country in the 1960s, there have been 13 outbreaks in addition to sporadic cases. An outbreak during 2000 coincided with the movement of sacrificial animals from rural to urban areas for the festival of Eid-ul-Azha. Diagnosis was suspected in patients with fever and thrombocytopenia, and confirmed retrospectively using immunoassays and reverse transcriptase-PCR. Patients were given platelet, plasma and red cell infusions. Management varied due to unfamiliarity with the condition and its treatment, lack of availability of diagnostic laboratory tests and limited supply of ribavirin. Inadequate antiviral treatment and late presentation probably contributed to the death of six of the eight patients. Renal failure, disseminated intravascular coagulation and persistent high-grade fever were associated with mortality. The nucleotide sequence of the small genomic RNA segment of the CCHFV isolated in this outbreak was found to be very closely related to the CCHFV strains previously isolated in Pakistan.
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PMID:Crimean-Congo hemorrhagic fever: experience at a tertiary care hospital in Karachi, Pakistan. 1593 14

In this article, the development of a new TaqMan-based one-step real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay for detection and quantification of Crimean-Congo hemorrhagic fever virus (CCHFV) RNA is described. Selected oligos targeting the highly conserved S region of CCHFV were designed by using our oligo design and analysis software, Oligoware 1.0. None of the primer sequences showed genomic cross-reactivity with other viruses or cells in a BLAST (NCBI) search analysis. The sensitivity and specificity of the primers and the probe were tested using 18 serum samples from patients from East Anatolian who were suspected of having CCHFV, including 2 samples that had already been confirmed to be positive for CCHFV. Among the 16 previously unconfirmed samples, 5 were positive by TaqMan-based one-step real-time RT-PCR and 1 was positive by non-nested RT-PCR, and these results were confirmed with DNA sequencing analysis. The 2 previously confirmed CCHFV RNA samples were also positive by both TaqMan-based one-step real-time RT-PCR and non-nested RT-PCR tests. To ensure the quantitative reproducibility of TaqMan-based one-step real-time RT-PCR, the procedure was repeated several times and the same results were obtained (SD = 0.84 [maximum value]). The developed assay was able to sensitively quantify the concentration of CCHFV RNA, which ranged from 10(2) to 10(7) copies/ml per reaction, using plasmid standards generated from the CCHFV RNA (correlation coefficiency = 0.989). The results of the one-step real-time RT-PCR assay were more sensitive than those of the non-nested RT-PCR assay. It can be concluded that our one-step real-time RT-PCR assay is a reliable, reproducible, specific, sensitive and simple tool for the detection and quantification of CCHFV.
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PMID:Rapid and quantitative detection of Crimean-Congo hemorrhagic fever virus by one-step real-time reverse transcriptase-PCR. 1637 67

Crimean-Congo haemorrhagic fever (CCHF) is an often fatal viral infection described in about 30 countries, and it has the most extensive geographic distribution of the medically important tickborne viral diseases, closely approximating the known global distribution of Hyalomma spp ticks. Human beings become infected through tick bites, by crushing infected ticks, after contact with a patient with CCHF during the acute phase of infection, or by contact with blood or tissues from viraemic livestock. Clinical features commonly show a dramatic progression characterised by haemorrhage, myalgia, and fever. The levels of liver enzymes, creatinine phosphokinase, and lactate dehydrogenase are raised, and bleeding markers are prolonged. Infection of the endothelium has a major pathogenic role. Besides direct infection of the endothelium, indirect damage by viral factors or virus-mediated host-derived soluble factors that cause endothelial activations and dysfunction are thought to occur. In diagnosis, enzyme-linked immunoassay and real-time reverse transcriptase PCR are used. Early diagnosis is critical for patient therapy and prevention of potential nosocomial infections. Supportive therapy is the most essential part of case management. Recent studies suggest that ribavirin is effective against CCHF, although definitive studies are not available. Health-care workers have a serious risk of infection, particularly during care of patients with haemorrhages from the nose, mouth, gums, vagina, and injection sites. Simple barrier precautions have been reported to be effective.
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PMID:Crimean-Congo haemorrhagic fever. 1655 45

Crimean-Congo haemorrhagic fever (CCHF) is potentially a fatal disease transmitted by tick bite, close contact with blood or tissues of infected humans or viraemic livestock. We present the clinical course of 2 breastfeeding women with CCHF and their babies. Both the mothers had positive reverse transcriptase-polymerase chain reaction for CCHF virus in blood and it was negative in breast milk. At follow-up, babies did not develop CCHF infection.
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PMID:Breastfeeding in Crimean-Congo haemorrhagic fever. 1793 82

Crimean-Congo haemorrhagic fever (CCHF) is a potentially fatal viral disease. In this study, the aim was to investigate the prognostic factors affecting the patient's survival and risk factors to fatality. At Ondokuz Mayis University Faculty of Medicine, a tertiary referral centre near the CCHF epidemic region, patients with typical clinical findings and indicative microbiological results for IgM and/or reverse transcriptase-polymerase chain reaction of CCHF virus were enrolled in the study, from 2004 to 2007. Patients were divided into two subgroups according to their survival outcomes; group I (n = 44) survived patients and group II (n = 6) consisted of fatal cases. The median platelet count was significantly lower in the fatal group (11000/mm(3)) when compared to the survived group (49500/mm(3)). Aspartate transferase and alanine transferase (ALT) levels were significantly higher in group II, when compared to group I. Also, the median range of serum lactic dehydrogenase (LDH) and creatinine phosphokinase (CPK) levels were much more elevated, and prothrombin time (PT) and activated partial thromboplastin time (aPTT) were prolonged in fatal cases. There was also a significant difference in median age of these two groups. Advanced age, late admission, low platelet count, increased AST, ALT, CPK and LDH levels, and prolonged PT and aPTT could be an early indicator of poor prognosis in patients with CCHF.
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PMID:Risk factors for fatality in patients with Crimean-Congo haemorrhagic fever. 1953 53

Crimean-Congo haemorrhagic fever (CCHF) is a viral haemorrhagic fever caused by the CCHF virus. It is mainly transmitted to humans and animals by ticks. In recent y, large numbers of livestock have been transported across the border areas of Ardabil Province resulting in an outbreak of CCHF in the adjacent districts. A comprehensive study was carried out to assess the epidemiological aspects of the disease in this province. In the study area, 130 ticks were collected from randomly selected villages and classified into 9 species of hard tick and 2 species of soft tick. All ticks were analyzed for the presence of CCHF virus genome using gel-based and real-time reverse transcriptase polymerase chain reactions (RT-PCR). The results showed CCHF infection in almost 28% of ticks collectively. Also, of 56 livestock sera, around 39% were IgG-positive. The presence of anti-CCHF virus IgG antibodies and the CCHF virus genome in ticks points to a great hidden threat of an outbreak in these districts. Those in high-risk professions in this province should be informed and trained on the risk of CCHF with urgency.
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PMID:A survey of Crimean-Congo haemorrhagic fever in livestock and ticks in Ardabil Province, Iran during 2004-2005. 1995 40

Crimean-Congo hemorrhagic fever (CCHF) is a fatal viral disease that occurs in approximately 30 countries. It has the most extensive geographic range among the tick-borne viruses that affect human health. Recently, a 6-year-old boy presented with complaints of fever, fatigue, and loss of appetite. He revealed a history of tick bite in rural Istanbul three days prior to presentation. A hyperemia was detected at the site of the tick bite. Laboratory tests showed that alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and creatine phosphokinase levels were elevated and that the prothrombin time and activated partial thromboplastin time were prolonged. Anti-CCHF virus IgM ELISA and a reverse transcriptase-PCR assay for CCHF RNA were both positive. Phylogenetic studies revealed that the virus was a new AP92-like CCHF strain, which was named KMAG-Hu-07-01 (accession number EU057975). This patient could provide important information on the transmission dynamics of CCHF infection.
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PMID:A newly identified Crimean-Congo hemorrhagic fever virus strain in Turkey. 2000 60


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