Detection Of Measles Virus Genome Research Articles

Phylogenetic and epidemiological analysis of measles outbreaks in Denmark, 2013 to 2014.

Despite the introduction of safe, effective vaccines decades ago and joint global public health efforts to eliminate measles, this vaccine-preventable disease continues to pose threats to children's health worldwide. During 2013 and 2014, measles virus was introduced into Denmark through several independent importations. This resulted in a number of secondary cases (n=7), with two clusters in 2013 and one in 2014. In total, there were 44 cases of measles. Most cases (n=41) were laboratory confirmed by detection of measles virus genome by real-time reverse transcription (RT)-PCR and IgM antibodies. The viruses from confirmed cases were genotyped by sequencing. Only one genotype circulated each year, i.e. D8 and B3, respectively. Sequencing of measles virus from different clinical specimens from the same patients revealed that sequence variants of measles viruses might co-exist and co-transmit during an outbreak. The majority of the cases were unvaccinated (n=27) or recipients of one dose of measles-mumps-rubella (MMR) vaccine (n=7). In addition, two fully vaccinated adult cases were reported in 2014. We demonstrate the transmission of measles virus in a population in which the two-dose MMR vaccination coverage rate was 80% and how even vaccinated individuals may be at risk of contracting measles once transmission has been established.

Application of a simple method for the detection of measles virus genome by loop-mediated isothermal amplification (LAMP)

A new simple RT-LAMP method was applied to detect measles virus nucleic acid and compared with nest-RT-PCR. Compare the detection rate of the RT-LAMP method with that of nest-RT-PCR by detecting measles virus nucleic acid from measles virus and clinical samples. The nucleic acid positive rates of all 23 strains of measles virus are all 100% by the two methods. But to the detection of 18 clinical samples which are negative in measles isolation, the nest-RT-LAMP showed 56.52% positive rate of nucleic acid of measles virus and nest-RT-PCR showed 47.83%. RT-LAMP is more sensitive than nest-RT-PCR.

A simple method for the detection of measles virus genome by loop-mediated isothermal amplification (LAMP)

Approximately 20,000–30,000 measles patients were reported in a surveillance of infectious diseases because of low vaccine coverage of 80% in Japan. Among them, some were thought to be secondary vaccine failure (SVF) with generally mild or non‐typical measles illness and sometimes became a source of further transmission. We have developed a new, sensitive, and rapid method to detect the measles virus genome by reverse transcription loop‐mediated isothermal amplification (RT‐LAMP). We examined 50 nasopharyngeal secretion (NPS) samples that were obtained during the 1999 outbreak and stored at −70°C and fresh NPS, lymphocytes and sera from 11 patients in 2003. Total RNA was extracted from the samples and subjected to reverse transcription‐polymerase chain reaction (RT‐PCR) and RT‐LAMP. We detected the genomic RNA corresponding to at least 0.01–0.04 TCID50, 30–100 copies in samples by RT‐LAMP within 60 min after extraction of RNA, and all four genotypes isolated in Japan were equally amplified. Specific DNA amplification was monitored spectrophotometrically by real time turbidimeter and the quantity of RNA was calculated. Measles virus genome was detected in 44 of 50 stored NPS by RT‐PCR and in 49 by RT‐LAMP. The vaccine strain was discriminated from wild strains after sequencing the LAMP products. RT‐LAMP is a useful rapid diagnostic method for the detection of measles virus without any special apparatus, showing higher sensitivity than RT‐PCR, and expected to be applied for hospital‐based infection control and for laboratory‐based measles surveillance. J. Med. Virol. 76:406–413, 2005. © 2005 Wiley‐Liss, Inc.

Detection of measles virus genome in bone-marrow aspirates from adults

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Detection of measles virus genome in bone-marrow aspirates from adults.

We investigated the presence of the measles virus genome in order to identify asymptomatic infections in the adult population. Bone-marrow aspirates were obtained from 179 patients, 20-96 years of age, for the diagnosis of malignant diseases (29 with malignant lymphoma, 28 with acute leukaemia, 21 with myelodysplastic syndrome, five with multiple myeloma and 96 with other diseases). The measles virus genome was detected in 17 (9.5%) of 179 individuals by RT-PCR and 28 (15.6%) through hybridization. The genomes detected in bone marrow were all in the same cluster, D5, the strain circulating during the study period, and no evidence of persistent infection was obtained. We conclude that asymptomatic infections of measles virus are common in adults and the presence of the measles virus genome would not be related to the pathogenesis of illness.

A case of intractable epilepsy positive for the detection of measles virus genome in the cerebrospinal fluid and peripheral mononuclear cells using reverse transcriptase-polymerase chain reaction

We report a rare case of intractable frontal lobe epilepsy with mental deterioration, in which the measles virus gene was detected from the cerebrospinal fluid (CSF) and peripheral mononuclear cells (PBMC) obtained 9 years after the first epileptic episode using reverse transcriptase-polymerase chain reaction (RT-PCR). The patient had been immunized with an attenuated measles vaccine and had no history of clinically apparent acute measles infection. However the analysis of the sequence of the PCR product from CSF showed the circulating wild strain genotype at the time when the patient complained of his first epileptic episode.

A case of intractable epilepsy with mental deterioration: Detection of measles virus genome in cerebrospinal fluid and peripheral mononuclear cells using reverse transcriptase‐polymerase chain reaction

Psychiatry and Clinical NeurosciencesVolume 49, Issue 3 p. S294-S295 A case of intractable epilepsy with mental deterioration: Detection of measles virus genome in cerebrospinal fluid and peripheral mononuclear cells using reverse transcriptase-polymerase chain reaction TASUKU MIYAJIMA MD, Corresponding Author TASUKU MIYAJIMA MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanDepartment of Pediatrics, Tokyo Medical College, 6–7-1 Nishi-shinjuku, Shinjuku-ku, Tokyo 160, Japan.Search for more papers by this authorHISASHI KAWASHIMA MD, HISASHI KAWASHIMA MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this authorAKINORI HOSHIKA MD, AKINORI HOSHIKA MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this authorMASAAKI OGIHARA MD, MASAAKI OGIHARA MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this authorNAOTO YAMADA MD, NAOTO YAMADA MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this authorWANG CHUAN-YU MD, WANG CHUAN-YU MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this authorKEIKO KINOUE MD, KEIKO KINOUE MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this authorKOUJI TAKEKUMA MD, KOUJI TAKEKUMA MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this author TASUKU MIYAJIMA MD, Corresponding Author TASUKU MIYAJIMA MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanDepartment of Pediatrics, Tokyo Medical College, 6–7-1 Nishi-shinjuku, Shinjuku-ku, Tokyo 160, Japan.Search for more papers by this authorHISASHI KAWASHIMA MD, HISASHI KAWASHIMA MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this authorAKINORI HOSHIKA MD, AKINORI HOSHIKA MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this authorMASAAKI OGIHARA MD, MASAAKI OGIHARA MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this authorNAOTO YAMADA MD, NAOTO YAMADA MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this authorWANG CHUAN-YU MD, WANG CHUAN-YU MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this authorKEIKO KINOUE MD, KEIKO KINOUE MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this authorKOUJI TAKEKUMA MD, KOUJI TAKEKUMA MD Department of Pediatrics, Tokyo Medical College, Tokyo, JapanSearch for more papers by this author First published: June 1995 https://doi.org/10.1111/j.1440-1819.1995.tb02214.xAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume49, Issue3June 1995Pages S294-S295 RelatedInformation

Detection of measles virus genome directly from clinical samples by reverse transcriptase-polymerase chain reaction and genetic variability

A simple and sensitive method for the detection of measles virus genome was developed, amplifying the regions encoding the nucleocapsid (N) protein and hemagglutinin (H) protein of measles virus by reverse transcriptase-polymerase chain reaction (RT-PCR). We examined a variety of measles patients: 28 patients with natural infection, 4 with measles encephalitis and 1 with subacute sclerosing panencephalitis (SSPE). In 28 patients with natural measles infection a single step PCR amplyfying the N region resulted in a high detection rate for all plasma samples (28/28) within 3 days of the onset of rash and 80% (20/25) even on day 7 of the onset of rash and later. Within 3 days of the onset of rash, 24/25 (96.0%) of nasopharyngeal secretions (NPS) and 27/28 (96.4%) of peripheral blood mononuclear cells (PBMC) were positive for the N region PCR and the positivity rate of PCR decreased in NPS and PBMC after 7 days of the rash. In acute measles infection, measles genome was detected in all cell fractions, CD4, CD8, B cells, and monocytes/ macrophages by the H gene nested PCR. Measles genome was also detected from cerebrospinal fluids (CSF) in patients with measles encephalitis, SSPE, and acute measles by the H gene nested PCR. PCR products of the N and H regions were sequenced and we confirmed the presence of measles genome. Based on the sequence data, chronological sequence differences were observed over the past 10 years. The sequences obtained from the SSPE patient were closely related to those of the wild viruses that were circulating at the time when the patient initially acquired measles. RT-PCR for NPS, PBMC, CSF, and plasma provides a useful method for the diagnosis of measles and molecular epidemiological study in addition to virus isolation.

Subacute Measles Encephalitis in the Young Immunocompromised Host: Report of Two Cases Diagnosed by Polymerase Chain Reaction and Treated with Ribavirin and Review of the Literature

Two young patients with subacute measles encephalitis are described: a 20-year-old male hemophiliac infected with human immunodeficiency virus (HIV) and a 4-year-old girl with acute leukemia. Both patients were afebrile and had persistent focal seizures and slurred speech beginning 2 and 7 months, respectively, after the onset of uncomplicated acute measles. The diagnosis of subacute measles encephalitis was established by demonstration of paramyxovirus nucleocapsid on electron microscopy of brain tissue in one case and by detection of measles virus genome with the polymerase chain reaction in both. Treatment of the HIV-infected man with intravenous ribavirin was begun when the patient lost consciousness after several weeks of seizures; he died. The girl with leukemia was treated early after the onset of symptoms and recovered after a 15-week course. Review of 31 previously published cases revealed a typical clinical presentation. Cerebrospinal fluid (CSF) analysis, electroencephalography, measurement of measles antibody in serum and CSF, and computed tomography of the brain were not helpful in the diagnosis of subacute measles encephalitis. In contrast, histologic examination of brain tissue proved useful in establishing the diagnosis. On the basis of our experience and our literature review, we conclude that histologic and polymerase chain reaction studies of brain tissue are required for the early diagnosis of subacute measles encephalitis and that therapy with intravenous ribavirin is effective when administered early.

Detection of measles virus genome in blood leucocytes of patients with certain autoimmune diseases.

RNA isolated from lymphocytes of peripheral blood was dot-hybridized to a hybrid plasmid containing specific sequences for measles virus nucleocapsid protein. Viral RNA was detected in the lymphocytes of 28 of 34 (82%) patients with systemic lupus erythematosus (SLE) and of 40 of 68 (59%) patients with chronic glomerulonephritis (CGN), and was not detected in 29 control patients.