Rabies virus detection by RT-PCR in decomposed naturally infected brains

David, Dan; Yakobson, B.; Rotenberg, Dorith; Dveres, N.; Davidson, Irit; Stram, Y.

doi:10.1016/s0378-1135(02)00041-x

Cited by 87 publications

(72 citation statements)

References 9 publications

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“…According to the World Health Organization (WHO) recommendations, the RT-PCR technique should not be used for routine post-mortem diagnosis of rabies if brain tissue is available; the direct fluorescent antibody test should be used [3]. However, in decomposed samples, where traditional methods lose their sensitivity, RT-PCR becomes an important diagnostic tool [7,10,11].…”

Section: Discussionmentioning

confidence: 99%

“…Using RT-PCR, David et al [11] found a positivity rate of 100%, but only after 36 days of exposure in large animals at tropical temperatures. Under natural conditions of decomposition, the size of the brains of these species could encourage the viral preservation given its greater mass in contrast to smaller animals such as mice, where the rabies virus was detected until day 15 using a heminested PCR assay [19].…”

Section: Discussionmentioning

confidence: 99%

“…Molecular biology techniques such as reverse transcriptase polymerase chain reaction (RT-PCR) are important diagnostic tools for the rabies virus [9][10][11][12][13]. They have also been used in several retrospective studies using samples stored under inadequate refrigeration for long periods of time [14].…”

Section: Introductionmentioning

confidence: 99%

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Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions

Beltrán¹,

Dohmen²,

Pietro

et al. 2014

J Infect Dev Ctries

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Introduction: The exposure of nervous tissue samples to high temperatures affects the sensitivity of rabies virus diagnostic tests, causing degradation of the viral structure. This study evaluated reverse transcriptase polymerase chain reaction (RT-PCR) for the diagnosis and molecular characterization of brain tissue samples in an advanced state of decomposition and poorly conserved viral isolates by comparing it with routine diagnostic tests. Methodology: A panel of three canine brain samples exposed to controlled decomposition for 7, 15, 30, and 120 days were evaluated using fluorescence antibody test (FAT), mouse inoculation test (MIT), and RT-PCR. In addition, 14 isolates of rabies variants, representing the largest circulation in Argentina, preserved in inadequate cooling for six to eight years were analyzed. Molecular typing of strains was performed using a 159-nucleotide region corresponding to the nucleoprotein gene. Results: The three samples analyzed were positive by RT-PCR at all the decomposition times evaluated, in contrast to results observed with FAT and MIT, which rapidly became negative. In addition, 100% of the inadequately preserved samples were characterized molecularly. The limit of detection of RT-PCR was 0.5 MICDL 50 /0.03 mL. Conclusion: RT-PCR can be useful for rabies diagnosis and typing of putrefying samples or rabies isolates stored in inadequate conditions.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions

Beltrán¹,

Dohmen²,

Pietro

et al. 2014

J Infect Dev Ctries

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“…Rapid Fluorescent Focus Inhibition Test (RFFIT) Amerika Birleßik DevletleriÕnde standart test olarak kullanÝlmaktadÝr (17). Son yÝl-larda Reverse Transcription-Polymerase Chain Reaction (RT-PCR) yšntemi, tanÝyÝ doÛrulamak i•in kullanÝlma-ya baßlanmÝß olup, šzellikle beyin dokusunun •ŸrŸmeye baßladÝÛÝ durumlarda kullanÝlabileceÛi gšsterilmißtir (5,18). †lkemizde de histopatolojik inceleme, FAT, RT-PCR ve FÜT yšntemleri kullanÝlmaktadÝr.…”

Section: Gürüþunclassified

Kuduz Hastalığına Bağlı Bir Adli Otopsi

Tuğcu¹,

Dalgıç²,

Öngürü³

et al. 2003

Bull Leg Med

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…ZETKuduz hastalÝÛÝ, Rhabdoviridea ailesinden LyssavirusÕun neden olduÛu, akut seyreden ve profilaksi yapÝlmadÝÛÝnda šlŸmle sonu•lanan viral bir enfeksiyondur. SaÛlÝk BakanlÝÛÝ verilerine gšre TŸrkiyeÕde 1973-2001 yÝllarÝ arasÝnda 529 kuduz hastalÝÛÝna baÛlÝ šlŸm olgusu bildirilmißtir. DŸnyadaki insan kuduz vakalarÝnÝn yÝllÝk insidansÝnÝn 25.000Õin Ÿzerinde olduÛu belirtilmektedir.Olgumuz, 21 yaßÝnda erkek olup bir kšpek tarafÝndan frontal bšlgesinden ÝsÝrÝldÝÛÝ, temas sonrasÝ profilaksinin yapÝldÝÛÝ, 25 gŸn sonra šldŸÛŸ bildirilmißtir.Kuduz hastalÝÛÝna baÛlÝ šlŸm olgularÝnda, kesin šlŸm nedeninin saptanabilmesi i•in genel otopsi uygulamalarÝ yanÝnda laboratuvar araßtÝrmalarÝn da yapÝlmasÝ gerekmektedir. Otopsi ekibinin, etkenin bulaßma riskine karßÝ šnlem almasÝ, otopsi salonunun ve otopsi aletlerinin dezenfeksiyonu ile postmortem inceleme i•in uygun dokular alÝnmasÝnda, ayrÝca uygun koßul-larda muhafaza edilerek laboratuvara gšnderilmesinde standart protokollerin uygulanmasÝ šnem taßÝmaktadÝr.Olgumuz, profilaksiye raÛmen šlŸme neden olabilecek faktšrlerin tartÝßÝlmasÝ ve kuduz hastalÝÛÝ ßŸpheli olgularÝn otopsilerinde dikkat edilmesi gereken šnlemlerin vurgulanmasÝ amacÝyla sunulmaktadÝr.Anahtar Kelimeler: Kuduz, otopsi, biyolojik korunma. SUMMARYRabies is an acute viral infectious disease with the highest mortality rate The causative agent is a neurotropic RNA virus in the family of Rhabdoviridae genus Lyssa virus. According to the records of Ministry of Health in Turkey; 529 people died from rabies between 1973-2001. It is estimated that human cases associated with rabies occur with an incidence of 25000 per year.Our case was a 21 years old male. He had a history of dog bite at the frontal site of his head and death occured within 25 days despite the prophylaxis.If a case is suspected to be dead because of infectious diseases, an autopsy can be performed without permission of relatives, by demand of officers, only.Spesific laboratory tests should also be done as a component of autopsy procedures in cases of human deaths associated with rabies. Autopsy team must be aware of the risk of transmission of virus and standard protocoles must be followed in disenfection of autopsy room, and instruments, and also in handling tissue samples.In this case, our purpose was to investigate and discuss the factors that may cause death in cases of rabies in spite of prophylaxis and highlight the biosafety considerations during autopsy in cases with suspicion of rabies.

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“…Classical reverse transcription-polymerase chain reaction (RT-PCR) assays on the other hand has been reported to be a sensitive and specific tool for routine diagnostic purposes [18,19]. These assays were also used to detect rabies virus in decomposed samples [1,6] and archival specimens [4,14,20]. Few of the brain tissue samples submitted for diagnosing rabies to our laboratory are often autolysed due to faulty transport conditions.…”

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confidence: 99%

Evaluation of RT-PCR Assay for Routine Laboratory Diagnosis of Rabies in Post Mortem Brain Samples from Different Species of Animals

et al. 2012

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Rabies in domestic and wild animals continues to be a major public health threat in India. Rapid and accurate diagnosis of rabies in animals is therefore of utmost importance as the individuals who were in contact with the rabid animals are at a greater risk. A significant amount of diagnostic tissue samples submitted to our laboratory are often autolysed and the WHO recommended direct fluorescent antibody test (FAT) for rabies diagnosis cannot be used in such samples. In this pilot study we have evaluated three different diagnostic primer sets for rapid sensitive and specific detection of rabies genome from the brain samples of different species of animals. We have validated a sensitive RT-PCR assay using brain tissue samples from different species of animals such as cat, cattle, dog, mouse and human, for routine diagnosis of rabies. Our results show the potential of this assay as a confirmatory test when the FAT results are unreliable and also as an alternative diagnostic test in circumstances when the diagnostic samples are unsuitable for use in FAT. Furthermore the nucleotide sequence of nucleoprotein gene amplified using this assay can also be used for the molecular epidemiological study of the rabies viruses in India.Keywords Rabies Á India Á Diagnosis Á FAT Á RT-PCR Animal and human rabies are endemic throughout the Indian subcontinent with the exception of Andaman and Nicobar Islands. The causative agent, rabies virus (family: Rhabdoviridae; genus: Lyssavirus) is largely maintained in two ecologically inter-related disease cycles; urban and sylvatic (wildlife). Although, dog is primarily responsible for rabies transmission to humans in India [3], wildlife plays a much lesser but not insignificant role in transmission [15]. It has been reported that more than 96 % of rabies incidence in India are the result of contact with infected dogs. In addition, rabies was reported as a result of contact with infected jackals in 1.7 %, cats in 0.8 %, monkeys in 0.4 %, mongooses in 0.4 %, and foxes in 3 % of the cases [3]. Therefore ante and post mortem diagnosis of rabies in animals is of utmost importance as the animals and human beings who come in contact with these rabid animals are at a greater risk of contracting the disease. The most widely used diagnostic test for rabies is the fluorescent antibody test (FAT), which is recommended by both WHO and OIE [17]. This 'gold standard' test may be used directly on a brain tissue impression smear and can also be used to confirm the presence of rabies antigen in cell culture or in brain tissue of mice that have been inoculated for diagnostic purposes. Although, the FAT gives reliable results on fresh specimens within a few hours in more than 95-99 % of the cases [17], autolysed tissue samples can reduce the sensitivity of this test and often are unsuitable for confirming the presence of rabies antigen. Classical reverse transcription-polymerase chain reaction (RT-PCR)

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Rabies virus detection by RT-PCR in decomposed naturally infected brains

Cited by 87 publications

References 9 publications

Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions

Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions

Kuduz Hastalığına Bağlı Bir Adli Otopsi

Evaluation of RT-PCR Assay for Routine Laboratory Diagnosis of Rabies in Post Mortem Brain Samples from Different Species of Animals

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