Prediction of the Spatial Origin of Puumala Virus Infections Using L Segment Sequences Derived from a Generic Screening PCR

To screen diagnostic specimens for the presence of hantavirus genomes or to identify new hantaviruses in nature, the pan-hanta L-PCR assay, a broadly reactive nested reverse transcription polymerase chain reaction (RT-PCR) assay targeting the L segment, is highly preferred over other assays because of its universality and high sensitivity. In contrast, the geographic allocation of Puumala virus strains to defined outbreak regions in Germany was previously done based on S segment sequences. We show that the routinely generated partial L segment sequences resulting from the pan-hanta L-PCR assay provide sufficient phylogenetic signal to inform the molecular epidemiology of the Puumala virus. Consequently, an additional S segment analysis seems no longer necessary for the identification of the spatial origin of a virus strain.

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Polymerase chain reaction amplification and typing of rotavirus in environmental samples

Water Science & Technology ◽

10.2166/wst.1995.0643 ◽

1995 ◽

Vol 31 (5-6) ◽

pp. 371-374 ◽

Cited By ~ 5

Author(s):

R. Gajardo ◽

R. M. Pintó ◽

A. Bosch

Keyword(s):

Polymerase Chain Reaction ◽

Environmental Samples ◽

Polymerase Chain Reaction Amplification ◽

Rt Pcr ◽

Chain Reaction ◽

Pcr Assay ◽

Group A ◽

Reaction Amplification ◽

Stool Specimens ◽

Polymerase Chain

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.

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Detection of multiple enteric virus strains within a foodborne outbreak of gastroenteritis: an indication of the source of contamination

Epidemiology and Infection ◽

10.1017/s0950268804003218 ◽

2004 ◽

Vol 133 (1) ◽

pp. 41-47 ◽

Cited By ~ 55

Author(s):

C. I. GALLIMORE ◽

C. PIPKIN ◽

H. SHRIMPTON ◽

A. D. GREEN ◽

Y. PICKFORD ◽

...

Keyword(s):

Arabian Gulf ◽

Enteric Virus ◽

Multiple Infections ◽

Rt Pcr ◽

Chain Reaction ◽

Epidemiological Analysis ◽

Viral Aetiology ◽

Polymerase Chain ◽

Most Probable Explanation ◽

Virus Strains

An outbreak of acute gastroenteritis of suspected viral aetiology occurred in April 2003 in the British Royal Fleet Auxillary ship (RFA) Argus deployed in the Northern Arabian Gulf. There were 37 cases amongst a crew of 400 personnel. Of 13 samples examined from cases amongst the crew, six enteric viruses were detected by reverse transcriptase polymerase chain reaction (RT–PCR). Five different viruses were identified including, three norovirus genotypes, a sapovirus and a rotavirus. No multiple infections were detected. A common food source was implicated in the outbreak and epidemiological analysis showed a statistically significant association with salad as the source of the outbreak, with a relative risk of 3·41 (95% confidence interval of 1·7–6·81) of eating salad on a particular date prior to the onset of symptoms. Faecal contamination of the salad at source was the most probable explanation for the diversity of viruses detected and characterized.

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BAALC and ERG Expression in Egyptian Patients with Acute Myeloid Leukemia, Relation to Survival and Response to Treatment

Open Access Macedonian Journal of Medical Sciences ◽

10.3889/oamjms.2016.058 ◽

2016 ◽

Vol 4 (2) ◽

pp. 264-270 ◽

Cited By ~ 2

Author(s):

Aml Soliman ◽

Asmaa Abdel Aal ◽

Reham Afify ◽

Noha Ibrahim

Keyword(s):

Polymerase Chain Reaction ◽

Acute Myeloid Leukemia ◽

Reverse Transcription ◽

Myeloid Leukemia ◽

Rt Pcr ◽

Chain Reaction ◽

Pcr Assay ◽

Polymerase Chain ◽

Age And Sex ◽

Acute Myeloid

AIM: Aim was to detect Brain and Acute Leukemia, Cytoplasmic (BAALC) and ETS-related gene (ERG) expression in patients with acute myeloid leukemia (AML) as well as to study their biologic and prognostic impact on the disease outcome and survival.PATIENTS AND METHODS: The current study was carried out on 44 patients with denovo acute myeloid leukemia, as well as 44 age and sex matched controls. The quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was performed for estimation of BAALC and ERG expression.RESULTS: The current study was carried out on 44 patients with denovo acute myeloid leukemia, as well as 44 age and sex matched controls. The quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was performed for estimation of BAALC and ERG expression. BAALC was expressed in 36 (81.82%) of AML cases versus 10 (22.72%) of the control group which was highly statistically significant (P < 0.001). While ERG was positive in 39(88.64%) of cases and 8(18.18 %) of controls and that was also highly statistically significant (P < 0.001).CONCLUSION: Further researches still needed to clarify the role of BAALC and ERG in the pathogenesis of leukemia and their importance as targets for treatment of AML.

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Multiplex Nested Reverse Transcription-Polymerase Chain Reaction for Respiratory Viruses in Acute Otitis Media

Annals of Otology Rhinology & Laryngology ◽

10.1177/000348940311200311 ◽

2003 ◽

Vol 112 (3) ◽

pp. 252-257 ◽

Cited By ~ 7

Author(s):

Toshio Ishibashi ◽

Hiroko Monobe ◽

Masanobu Shinogami ◽

Yuka Nomura ◽

Jun Yano

Keyword(s):

Polymerase Chain Reaction ◽

Otitis Media ◽

Reverse Transcription ◽

Acute Otitis Media ◽

Respiratory Viruses ◽

Molecular Technique ◽

Rt Pcr ◽

Chain Reaction ◽

Pcr Assay ◽

Polymerase Chain

Because respiratory viruses play an important role in the causation and pathogenesis of acute otitis media (AOM), determining which virus has infected a child is important with respect to vaccines and antiviral drugs. In some instances, this information might be used to prevent the occurrence of AOM. We used a rapid, economical, and sensitive diagnostic system involving a multiplex nested reverse transcription–polymerase chain reaction (RT-PCR) assay to detect various respiratory viruses in clinical specimens of middle ear fluid (MEF) from children with AOM in our hospital. Multiplex RT-PCR was completed on 40 MEF samples from 28 infants and children less than 6 years old with AOM. Viral RNA was detected in 17 MEF samples (43%). Respiratory syncytial virus type A was present in 12 samples, adenovirus in 3, rhinovirus in 2, and influenza A (H3N2) in 1. The multiplex RT-PCR assay is recommended to clinical laboratories that are considering adoption of a molecular technique for viral diagnosis.

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Collaborative Trial for Validation of a Real-Time Reverse Transcriptase-Polymerase Chain Reaction Assay for Detection of Central Nervous System Tissues as Bovine Spongiform Encephalopathy RiskMaterial: Part 1

Journal of AOAC International ◽

10.1093/jaoac/89.5.1335 ◽

2006 ◽

Vol 89 (5) ◽

pp. 1335-1340

Author(s):

Amir Abdulmawjood ◽

Holger Schnenbrcher ◽

Michael BÜlte

Keyword(s):

Central Nervous System ◽

Polymerase Chain Reaction ◽

Real Time ◽

Meat Products ◽

Spongiform Encephalopathy ◽

Rt Pcr ◽

Chain Reaction ◽

Pcr Assay ◽

Heat Treated ◽

Polymerase Chain

Abstract A collaborative trial was conducted to evaluate a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay for detection of central nervous system (CNS) tissues in meat products (e.g., sausages). The method is based on the detection of ruminant glial fibrillary acidic protein (GFAP) mRNA by applying real-time RT-PCR. The assay was evaluated through a multicenter trial involving 12 participating laboratories that received coded cDNA obtained from 3 different types of sausages. The participants used 5 different real-time detection systems. The results obtained in this validation revealed that this real-time RT-PCR assay performed well in the different laboratories with a detection limit of at least 0.1% CNS in those test materials that contained strongly heat-treated samples (sausages cooked at 120C) and the medium heat-treated samples (sausages cooked at 80C). The detection limit of liver sausages was determined to be 0.2% of CNS. Neither the samples with no CNS additive nor the bovine DNA and the negative control containing 100% swine brain gave any positive signals. The presented results indicate that the real-time RT-PCR assay was just as reproducible between laboratories, as repeatable within a laboratory, could reliably be used for detection of bovine spongiform encephalopathy risk material in meat and meat products, and signify that it may be used with confidence in any laboratory.

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Detection of Occult Melanoma Cells in Paraffin-Embedded Histologically Negative Sentinel Lymph Nodes Using a Reverse Transcriptase Polymerase Chain Reaction Assay

Journal of Clinical Oncology ◽

10.1200/jco.2001.19.5.1437 ◽

2001 ◽

Vol 19 (5) ◽

pp. 1437-1443 ◽

Cited By ~ 50

Author(s):

Giuseppe Palmieri ◽

Paolo A. Ascierto ◽

Antonio Cossu ◽

Nicola Mozzillo ◽

Maria L. Motti ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Reverse Transcriptase ◽

Prognostic Significance ◽

Primary Melanoma ◽

Disease Stage ◽

Occult Metastasis ◽

Rt Pcr ◽

Chain Reaction ◽

Pcr Assay ◽

Polymerase Chain

PURPOSE: Detection of occult metastasis before the development of clinical disease could allow more accurate staging, appropriate follow-up procedures, and adjuvant therapies in patients with malignant melanoma (MM). The sentinel lymph node (SLN) has been proposed as a reliable predictor of metastatic disease in the lymphatic basin draining the primary melanoma. In this study, we screened both paraffin-embedded SLNs and peripheral-blood (PB) samples from MM patients at various stage of disease using a multimarker reverse transcriptase polymerase chain reaction (RT-PCR) assay. The prognostic significance of the presence of PCR-positive markers was also evaluated. PATIENTS AND METHODS: Total RNA was obtained from paraffin-embedded SLN sections and PB samples of 75 MM patients. RT-PCR was performed using tyrosinase and MelanA/MART1 as melanoma-associated markers. Radiolabeled PCR products were analyzed on denaturing polyacrylamide gels. RESULTS: Good sensitivity of the RT-PCR assay on archival tissues was demonstrated after comparison of RT-PCR results on frozen and paraffin-embedded SLNs from 16 MM patients. Significant correlation between the disease stage and marker expression in both PB and SLN samples was observed; the highest value was for patients who were positive for both markers in SLN (P = .006). Progression of disease was significantly associated with the total number of PCR-positive markers in both PB (P = .034) and SLN (P = .001) samples. CONCLUSION: Although sensitivity is lowered by the use of paraffin-embedded specimens, our data indicate that RT-PCR analysis of serial sections from archival SLNs may be helpful in improving detection of occult micrometastases, thus improving staging of patients with melanoma.

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Comparison of Virus Isolation and Reverse Transcription Polymerase Chain Reaction Assay for Detection of Bovine Viral Diarrhea Virus in Bulk Milk Tank Samples

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063870001200219 ◽

2000 ◽

Vol 12 (2) ◽

pp. 184-186 ◽

Cited By ~ 41

Author(s):

R. W. Renshaw ◽

R. Ray ◽

E. J. Dubovi

Keyword(s):

Infected Animal ◽

Virus Isolation ◽

Rt Pcr ◽

Chain Reaction ◽

Pcr Assay ◽

Diarrhea Virus ◽

Bulk Milk ◽

Polymerase Chain ◽

Viral Diarrhea Virus ◽

Bulk Milk Tank

The use of a reverse transcriptase polymerase chain reaction (RT-PCR) assay to screen bulk milk tank samples for bovine viral diarrhea virus (BVDV) has proven to be a sensitive and economical means to evaluate the lactating animals in a herd. The assay is capable of detecting the presence of a single persistently infected animal within a group of several hundred cows. Over a 3–year period, 144 samples from 97 farms were tested for BVDV using an RT-PCR assay in conjunction with a classical virus isolation (VI) procedure to measure the relative effectiveness of the techniques. Virus could be detected with both methods when the milk from a single persistently infected animal was diluted 1:600 with the milk from a herd of BVDV-negative animals. Based on individual farms, there was an overall prevalence of 12.4% BVDV infection, and the correlation between the 2 assays was 95.9%. In terms of sensitivity, specificity, and turnaround time, RT-PCR was superior to VI. However, of the 17 samples that were VI positive, 4 were RT-PCR negative. RT-PCR may not detect all naturally occurring BVDV isolates because they may contain minor sequence variations in the primer regions. VI and RT-PCR are both suitable for detection of BVDV in bulk milk samples when used independently, but to increase the probability of successful detection and to provide cross-checks against assay contamination, it is desirable to utilize both methods in parallel.

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