Applications of a duplex reverse transcription polymerase chain reaction and direct immunofluorescence assay in comparison with virus isolation for detection of influenza A and B

Abstract Background Persistence of Zika virus (ZIKV) ribonucleic acid (RNA) in semen is common after infection. Methods We designed a reverse-transcription polymerase chain reaction assay that targets antisense ZIKV RNA (asRNA) to assess ZIKV replication competence in ZIKV RNA-positive semen samples. Results We detected ZIKV asRNA in semen of 9 of 19 men (47.4%) diagnosed with ZIKV infection. All asRNA-positive samples had high ZIKV loads (cycle threshold values <26) and were obtained within 21 days of symptom onset. Conclusions The sensitivity of the asRNA assay for detection of ZIKV replication was higher than that of conventional virus isolation methods (47.4% vs 21.1%, P = .032).

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Corrigendum to: Relative Frequency of Seasonal Influenza A and B in Khuzestan Province by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) During 2009-2010 [published in Jundishapur J Microbiol. 2013;6(4):e5312]

Jundishapur Journal of Microbiology ◽

10.5812/jjm.16587 ◽

2013 ◽

Vol 6 (10) ◽

Author(s):

Zamaneh Hajikhezri ◽

Manoochehr Makvndi ◽

Ali Reza Samarbaf-zadeh ◽

Niloofar Neisi ◽

Kambiz Ahmadi

Keyword(s):

Polymerase Chain Reaction ◽

Reverse Transcription ◽

Relative Frequency ◽

Influenza A ◽

Seasonal Influenza ◽

Rt Pcr ◽

Chain Reaction ◽

Khuzestan Province ◽

Polymerase Chain

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Evaluation of a 384–Well Format for High-Throughput Real-Time Reverse Transcription Polymerase Chain Reaction for Avian Influenza Testing

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063870902100513 ◽

2009 ◽

Vol 21 (5) ◽

pp. 679-683 ◽

Cited By ~ 4

Author(s):

Pamela J. Ferro ◽

Jason Osterstock ◽

Bo Norby ◽

Geoffrey T. Fosgate ◽

Blanca Lupiani

Keyword(s):

Polymerase Chain Reaction ◽

Avian Influenza ◽

Real Time ◽

High Throughput ◽

Reverse Transcription ◽

Virus Isolation ◽

Rt Pcr ◽

Chain Reaction ◽

Response Planning ◽

Polymerase Chain

As concerns over the global spread of highly pathogenic avian influenza H5N1 have heightened, more countries are faced with increased surveillance efforts and incident response planning for handling a potential outbreak. The incorporation of molecular techniques in most diagnostic laboratories has enabled fast and efficient testing of many agents of concern, including avian influenza. However, the need for high-throughput testing remains. In this study, the use of a 384–well format for high-throughput real-time reverse transcription polymerase chain reaction (real-time RT-PCR) testing for avian influenza is described. The analytical sensitivity of a real-time RT-PCR assay for avian influenza virus matrix gene with the use of both 96– and 384–well assay formats and serial dilutions of transcribed control RNA were comparable, resulting in similar limits of detection. Of 28 hunter-collected cloacal swabs that were positive by virus isolation, 26 (92.9%) and 27 (96.4%) were positive in the 96– and 384–well assays, respectively; of the 340 hunter-collected swabs that were negative by virus isolation, 45 (13.2%) and 23 (6.8%) were positive in the 96– and 384–well assays, respectively. The data presented herein supports the utility of the 384–well format in the event of an avian influenza outbreak for high-throughput real-time RT-PCR testing.

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