Measles virus TaqMan RT-PCR (F gene; no longer in regular use; see Guidelines) v3 (protocols.io.8b9hsr6)

SUMMARYThe purpose of this work was the molecular study of the virus strain that caused the last measles outbreak in Greece. Twenty-four saliva specimens were obtained from selected patients serologically confirmed as measles cases between December 2005 and March 2006. Measles virus (MV) detection was performed by a nested RT–PCR. The 447-bp segment of the N gene of these MV strains was used for genotyping. The N gene sequences of the Greek MV strains were identical to each other, therefore a phylogenetic tree was constructed using one representative MV (ThesGRE/06). Our data show that the MV strain which caused the 2005–2006 outbreak in Greece belongs to genotype D6, and differs by 0·68% from the New Jersey D6 strain and by 5·5% from the MV vaccine strain Edmonston B (U03656) belonging to genotype A.

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

Journal of General Virology ◽

10.1099/0022-1317-83-10-2485 ◽

2002 ◽

Vol 83 (10) ◽

pp. 2485-2488 ◽

Cited By ~ 8

Author(s):

Satomi Sonoda ◽

Mitsuo Kitahara ◽

Tetsuo Nakayama

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Myelodysplastic Syndrome ◽

Persistent Infection ◽

Measles Virus ◽

Adult Population ◽

Virus Genome ◽

Malignant Diseases ◽

Rt Pcr ◽

Asymptomatic Infections

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.

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P1444 Evaluation of a real-time RT-PCR assay for the diagnosis of measles virus infections during a measles outbreak in Greece

International Journal of Antimicrobial Agents ◽

10.1016/s0924-8579(07)71283-9 ◽

2007 ◽

Vol 29 ◽

pp. S403

Author(s):

S. Kokotas ◽

M. Giannaki ◽

E. Horefti ◽

D. Sgouras ◽

M. Logothetis ◽

...

Keyword(s):

Real Time ◽

Measles Virus ◽

Virus Infections ◽

Rt Pcr ◽

Pcr Assay ◽

Measles Outbreak

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Rapid Identification of Measles Virus Vaccine Genotype by Real-Time PCR

Journal of Clinical Microbiology ◽

10.1128/jcm.01879-16 ◽

2016 ◽

Vol 55 (3) ◽

pp. 735-743 ◽

Cited By ~ 13

Author(s):

Felicia Roy ◽

Lillian Mendoza ◽

Joanne Hiebert ◽

Rebecca J. McNall ◽

Bettina Bankamp ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Measles Virus ◽

Rapid Identification ◽

Rt Pcr ◽

Outbreak Response ◽

Reverse Transcription Pcr ◽

Pcr Method ◽

Measles Outbreaks ◽

Genotype A

ABSTRACT During measles outbreaks, it is important to be able to rapidly distinguish between measles cases and vaccine reactions to avoid unnecessary outbreak response measures such as case isolation and contact investigations. We have developed a real-time reverse transcription-PCR (RT-PCR) method specific for genotype A measles virus (MeV) (MeVA RT-quantitative PCR [RT-qPCR]) that can identify measles vaccine strains rapidly, with high throughput, and without the need for sequencing to determine the genotype. We have evaluated the method independently in three measles reference laboratories using two platforms, the Roche LightCycler 480 system and the Applied Biosystems (ABI) 7500 real-time PCR system. In comparison to the standard real-time RT-PCR method, the MeVA RT-qPCR showed 99.5% specificity for genotype A and 94% sensitivity for both platforms. The new assay was able to detect RNA from five currently used vaccine strains, AIK-C, CAM-70, Edmonston-Zagreb, Moraten, and Shanghai-191. The MeVA RT-qPCR assay has been used successfully for measles surveillance in reference laboratories, and it could be readily deployed to national and subnational laboratories on a wide scale.

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Is the Measles Virus Indeed Involved in Carcinogenesis? – Commentary

International Journal of Negative Results ◽

10.14302/issn.2641-9181.ijnr-18-2195 ◽

2018 ◽

Vol 1 (1) ◽

pp. 20-23

Author(s):

Daniel Benharroch

Keyword(s):

Hodgkin Lymphoma ◽

Measles Virus ◽

Rt Pcr ◽

Beer Sheva ◽

Virus Rna ◽

Classic Hodgkin Lymphoma ◽

Repeated Calls ◽

Measles Antibodies ◽

International Experts

Objectives: An association between the measles virus and Hodgkin lymphoma has been disclosed by our laboratory in Beer-Sheva, starting in 2003. We question the refutation of our study and the absence of interest among experts. Methodology: It was based on immunohistochemistry with commercial, as well as experimental anti-measles antibodies. It relied also on RT-PCR and in situ hybridization evidence of measles virus RNA. Key Results: At this stage (2004), the link between the virus and the lymphoma was essentially descriptive. The first and last response to our challenge appeared in 2007, in the form of doublet articles, in the same issue of a major cancer journal. The two European research groups responding, rejected categorically our findings by proposing different arguments. Major Conclusion: As reservations to these reactions became soon apparent, a series of papers from our laboratory were published. These articles concerned the evidence of a relationship between the measles virus and additional categories of cancers. Different malignancies in which this virus was not expressed at all, were also described. A further study suggested a mechanism by which the measles virus may activate lymphomagenesis in classic Hodgkin lymphoma. To our dismay, and in spite of repeated calls to verify the various results, no further response was obtained from international experts.

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

Eurosurveillance ◽

10.2807/1560-7917.es.2015.20.39.30027 ◽

2015 ◽

Vol 20 (39) ◽

Cited By ~ 6

Author(s):

Lasse Dam Rasmussen ◽

Jannik Fonager ◽

Lisbet Krause Knudsen ◽

Peter Henrik Senten Andersen ◽

Jesper Rønn ◽

...

Keyword(s):

Measles Virus ◽

Virus Genome ◽

Vaccine Preventable Disease ◽

Global Public Health ◽

Rt Pcr ◽

Mmr Vaccine ◽

Epidemiological Analysis ◽

Clinical Specimens ◽

Vaccination Coverage Rate ◽

Measles Outbreaks

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.

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Pathotyping of Newcastle disease virus: A novel single BsaHI digestion method of detection and differentiation of avirulent strains (lentogenic and mesogenic vaccine strains) from virulent virus

10.1101/2021.07.16.452760 ◽

2021 ◽

Author(s):

Perumal Arumugam Desingu ◽

Shambhu Dayal Singh ◽

Kuldeep Dhama ◽

Obli Rajendran Vinodhkumar ◽

K Nagarajan ◽

...

Keyword(s):

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Rt Pcr ◽

Specific Primer ◽

Digestion Method ◽

F Gene ◽

Virulent Strains ◽

Virulent Virus ◽

Short Space

We provide a novel single restriction enzyme (RE) (BsaHI) digestion approach for detecting distinct pathotypes of the Newcastle disease virus (NDV). After scanning 4000 F gene nucleotide sequences in the NCBI database, a single RE (BsaHI) digesting site was discovered in the cleavage site. APMV-I "F gene" Class II specific primer-based reverse transcriptase PCR (RT-PCR) was utilized to amplify a 535 bp fragment, which was then digested with a single RE (BsaHI) for pathotyping avian NDV field isolates and pigeon paramyxovirus-1 isolates. The avirulent (lentogenic and mesogenic strains) produce 189 and 346 bp fragments, respectively, but the result in velogenic strains remains undigested with 535 bp fragments. In addition, 45 field NDV isolates and 8 vaccine strains were used to confirm the approach. The sequence-based analysis also agrees with the data obtained utilizing the single RE (BsaHI) digestion approach. The proposed technique had the potential to distinguish between avirulent and virulent strains in a short space of time, making it valuable in NDV surveillance and monitoring research.

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Persistent Measles Virus Infection and Otosclerosis

Annals of Otology Rhinology & Laryngology ◽

10.1177/000348940111001001 ◽

2001 ◽

Vol 110 (10) ◽

pp. 897-903 ◽

Cited By ~ 22

Author(s):

Wolfgang Arnold ◽

Hans P. Niedermeyer ◽

Maria Schuster ◽

Wolfgang J. Neubert ◽

Christa Baumann ◽

...

Keyword(s):

Cell Culture ◽

Polymerase Chain Reaction ◽

Measles Virus ◽

Otic Capsule ◽

Rt Pcr ◽

Chain Reaction ◽

Virus Rna ◽

Polymerase Chain ◽

H Protein ◽

Antibody Pattern

The cause of otosclerosis is still unknown. Recently, measles virus involvement has been implicated. The aim of this study was to analyze the presence of measles virus RNA within the otosclerotic focus and to evaluate the perilymphatic antibody pattern. Bone and perilymph specimens from 40 patients with the spontaneous form of otosclerosis and from control patients were investigated by reverse transcription polymerase chain reaction (RT-PCR), Western blot techniques, and cell culture. By the use of RT-PCR, measles virus RNA could be detected in 32 patients, but not in controls. Analysis of perilymph revealed the presence of antibodies to N, F1, and M measles virus proteins in all cases, and antibodies against H protein in 2 additional cases. In preosteoblasts cultured from otosclerotic bone chips, no measles virus RNA could be amplified. We conclude that the spontaneous form of otosclerosis is, in the vast majority of cases, a measles virus-associated disease of the otic capsule.

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Molecular Epidemiological Investigation of Human Parainfluenza 3 Virus Outbreak in a Pediatric Bone Marrow Transplantation Unit

Open Forum Infectious Diseases ◽

10.1093/ofid/ofx163.869 ◽

2017 ◽

Vol 4 (suppl_1) ◽

pp. S358-S359

Author(s):

Patrick Stapleton ◽

Stephen Perusini ◽

Angela Thomas ◽

Michelle Science ◽

Tal Schechter-Finkelstein ◽

...

Keyword(s):

Bone Marrow ◽

Phylogenetic Analysis ◽

Molecular Typing ◽

Gene Sequence ◽

Likelihood Method ◽

Positive Pressure ◽

Rt Pcr ◽

Hematopoietic Stem ◽

F Gene ◽

Hn Gene

Abstract Background Human parainfluenza virus 3 (hPIV3), a common cause of respiratory infections in children, can cause nosocomial outbreaks in patients undergoing hematopoietic stem cell transplantation, resulting in significant morbidity and mortality. Between July and August 2016, an increased number of hPIV3 infections were noted in a pediatric bone marrow transplant unit (BMT). Two patients were identified in late July and 4 patients in August. We undertook molecular typing of hPIV3 to determine whether cases represented multiple introductions of community virus strains or patient to patient transmission of a single strain. Previous reports of molecular typing have targeted either the F (fusion protein) gene or HN (hemagglutinin-neuraminidase) gene. We compared results using both methods direct from clinical specimens. Methods Nasopharyngeal (NP) swabs from 6 patients in BMT ward and 6 patients hospitalized on other wards had hPIV3 detected by the Luminex NxTAG Respiratory Pathogen Panel over 2 months. For the F gene a single pair of primers were used to first amplify then sequence a 278 basepair (bp) region by reverse-transcriptase PCR (RT-PCR). For HN gene a 1719 bp region was amplified using nested RT-PCR, then sequenced with 6 sets of overlapping primers. The resulting contigs were assembled manually with ContigExpress. Phylogenetic analysis of assembled sequences was performed in MEGA7 using the maximum likelihood method. Results For the HN gene sequence of 1715 bp was obtained for 10 of 12 patients (5 in each group). Phylogenetic analysis of HN sequences indicated 2 distinct hPIV3 lineages (Figure 1). The 5 BMT patients differed by a maximum of 1bp, while 5 samples from other wards differed by 14 to 57 bp. For the F gene only 98 bp of common sequence was obtained for 7 patients, all of whom had HN gene sequences available. Phylogenetic analysis of F gene sequence also supported the presence of 2 distinct lineages. Conclusion Molecular typing of hPIV3 suggests there was transmission of a single hPIV3 strain within the BMT unit despite protective isolation of all BMT patients in positive pressure single rooms and the use of contact and droplet precautions for infected cases. We found sequencing the HN gene more informative than sequencing the F gene. Disclosures All authors: No reported disclosures.

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Measles resurgence in Argentina: 1997–8 outbreak

Epidemiology and Infection ◽

10.1017/s0950268899003659 ◽

2000 ◽

Vol 124 (2) ◽

pp. 289-293 ◽

Cited By ~ 6

Author(s):

M. D. BILKIS ◽

P. R. BARRERO ◽

A. S. MISTCHENKO

Keyword(s):

Intensive Care ◽

Molecular Epidemiology ◽

Measles Virus ◽

Buenos Aires ◽

Clinical Findings ◽

Hospitalized Children ◽

Rt Pcr ◽

Sensitive Tool ◽

Catch Up

Epidemiological and clinical findings from 1162 serologically confirmed measles cases occurring in Buenos Aires, Argentina in 1997 and 1998 were retrospectively reviewed. From 90 hospitalized children, measles virus was detected by direct RT–PCR from nasopharyngeal secretions. Patients were grouped as follows: (i) not vaccinated: infants < 12 months; (ii) regularly vaccinated: children 1–4 years not covered by the last catch-up; (iii) catch-up vaccinated: patients 5–19 years immunized during the 1993 campaign. Most cases were recorded in non-vaccinated infants (54%), and the lowest in catch-up vaccinated children (16%). Mean age of the 90 hospitalized children was 11·3 months. Pneumonia was the major hospitalization cause followed by pneumonitis. Two children required intensive care and one died. The 1993 catch-up campaign seemed to reduce the number of cases in the 5- to 19-year-old group. Lack of timely follow-up probably led to the accumulation of susceptible individuals allowing measles re-emergence. Direct viral detection by RT–PCR proved to be a sensitive tool for molecular epidemiology surveillance.

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