Abstract
Aim: Early diagnosis of dreadful rabies is of utmost importance to restrict number of contacts and timely administration of post exposure prophylaxis. The present study was conducted to evaluate the sensitivity comparison of Nested RT-PCR with TaqMan real time PCR technique for intravitam diagnosis of rabies in animals from urine samples. Materials and Methods: Advance molecular approaches Nested RT-PCR and TaqMan real time PCR was employed on 21 urine samples for intravitam diagnosis of rabies. Comparison of both the techniques was done with standard immunofluorescence test (FAT) applied on brain for postmortem confirmation of rabies. Result: Rabies viral RNA was detected in 6/21 (28.57%) and 11/21 (52.38%) urine samples by application of Nested RT-PCR and TaqMan real time PCR respectively. Sensitivity obtained from both the techniques was 62.50% and 78.94% respectively when compared with gold standard immunofluorescence test (FAT). Conclusion: TaqMan real time PCR can serve as more sensitive and viable approach for the intravitam diagnosis of rabies as compared to Nested RT-PCR for detection of rabies from urine of suspected animals. Clinical importance: This study may serve as background for future intravitam rabies diagnostics.
Highlights
Since time immemorial rabies continues to be a major health threat to mankind as well as all warm blooded animals
In TaqMan real time PCR samples in which threshold cycle number (Ct) values were found to be in the range of 20-35 were considered positive and above 35 were considered negative [14]
With this technique viral RNA could be diagnosed in 11/21 (52.38%) urine samples sensitivity of 78.94% was obtained when compared with gold standard immunofluorescence test (FAT) on brain (Table-3)
Summary
Since time immemorial rabies continues to be a major health threat to mankind as well as all warm blooded animals. According to results of global surveillance by the World Health Organization, about 50,000 cases of human rabies occur each year [1], the majority of them in developing countries [2]. In India rabies is enzootic and is a serious public health and economic problem [3]. The appearance of specific rabies disease symptoms is preceded by prodromal period in which there are a number of non-specific symptoms of malaise [4]. With the advent of molecular approaches, it is possible to detect rabies ante-mortem from range of biological samples e.g. nuchal skin biopsy [6], saliva [7], Cerebrospinal fluid (CSF) [8] and urine samples [9]
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