Diagnosis Of Human Rabies Research Articles

A new choice for human rabies diagnosis: A case report of metagenomics next-generation sequencing in diagnosis of human rabies

PurposeWe report a case of human rabies diagnosed by the metagenomics next-generation sequencing (mNGS). A 59-year-old man developed clinical rabies 20 days after he was bitten by dogs. Treatment included induction of coma initially; rabies vaccine was not administered. The patient was treated with propofol, midazolam, recombinant human interferon α2b, ribavirin, and amantadine. Penehyclidine was administrated to relieved the rabies induced pulmonary edema and the salivation. ResultsThe patient’s situation got worse on the 26th day after admition, and died on the 29th day finally. ConclusionThe mNGS might be a new choice for human rabies diagnosis，penehyclidine was effective in decreasing the rabies induced pulmonary edema and the salivation.

Human rabies in China: evidence-based suggestions for improved case detection and data gathering

BackgroundChina still suffers heavily from rabies, although reported human cases continue to decrease year over year. There are far fewer laboratory-confirmed human cases than clinically diagnosed cases, which is a big problem that needs to be addressed. In this report, we summarize analyses of all specimens from human cases tested in our laboratory over the past 15 years, in order to promote laboratory diagnosis of rabies.MethodsFrom 2005 to 2019, a total of 271 samples from 164 suspected rabies cases were collected from local hospitals by the local Centers for Disease Control and Prevention (CDCs) in China. Saliva, cerebrospinal fluid (CSF), serum (blood) and urine were collected for ante-mortem diagnosis, and brain tissue, neck skin tissue and cornea were collected for post-mortem diagnosis. All of the specimens were tested by reverse transcription-polymerase chain reaction (RT-PCR), and brain tissues were also tested using fluorescent antibody test (FAT). The number of positive test results obtained using different fluids or tissues, and at different stages of the disease, were compared using a chi-square test and a more effective sampling program is recommended.ResultsAs the national reference laboratory for rabies surveillance in China, our laboratory has tested 271 samples from 164 suspected rabies cases collected by local CDCs since 2005. We found that saliva gave the highest number of positive test results (32%), compared with CSF and other fluids. We also found that serum or blood specimens collected in the last 3 days of life can test positive by RT-PCR.ConclusionsSerum or blood samples collected in the last 3 days of a patient’s life can be used to measure viral RNA, which means that serum samples, as well as saliva and CSF, can be used to detect viral RNA for anti-mortem diagnosis of rabies. Because of our findings, we have modified our “National Surveillance Project for Human Rabies”, by adding the collection and testing of serum samples from the end of the survival period. This will improve our national surveillance and laboratory diagnosis of human rabies.

Utility of forensic detection of rabies virus in decomposed exhumed dog carcasses

This report describes four suspected rabies cases in domestic dogs that were involved in human exposures. In all these cases, the animals were buried for substantial times before rabies testing was performed. Animal rabies is endemic in South Africa and domestic dogs are the main vector for transmission to humans. Diagnosis of rabies in humans is complicated, and diagnosis in the animal vector can provide circumstantial evidence to support clinical diagnosis of rabies in humans. The gold standard diagnostic method, fluorescent antibody test (FAT), only delivers reliable results when performed on fresh brain material and therefore decomposed samples are rarely submitted for diagnostic testing. Severely decomposed brain material was tested for the presence of rabies virus genomic material using a quantitative real-time reverse transcription polymerase chain reaction (q-real-time RT-PCR) assay when conventional molecular methods were unsuccessful. This may be a useful tool in the investigation of cases where the opportunity to sample the suspected animals post mortem was forfeited and which would not be possible with conventional testing methodologies because of the decomposition of the material.

Presence of Cross-Reactions with Other Viral Encephalitides in the Indirect Fluorescent-Antibody Test for Diagnosis of Rabies

The antemortem diagnosis of rabies in humans employs techniques that require accuracy, speed, and sensitivity. A combination of histochemical analysis, in vitro virus isolation, immunological methods, and molecular amplification procedures are utilized in efforts to diagnose the disease. Modern medicine now offers potentially life-saving treatment for a disease that was considered invariably fatal once clinical signs develop. However, medical intervention efforts require a rapid and accurate diagnosis as early in the course of clinical disease as possible. Indirect fluorescent-antibody (IFA) testing on cerebrospinal fluid and serum specimens provides rapid results, but the specificity of the assay has not been well studied. Because false-positive IFA results could significantly affect patient treatment and outcomes, it is critical to understand the specificity of this assay. In this study, IFA testing was performed on 135 cerebrospinal fluid and serum specimens taken from patients with viral encephalitis or a presumed viral infection involving an agent other than rabies virus. Results indicate that false-positive results can occur in interpreting the rabies IFA test. Staining patterns morphologically similar to antirabies staining were observed in 7 of the 135 cerebrospinal fluid specimens examined. In addition, a majority of the cerebrospinal fluid specimens tested from patients with encephalitis presented immunoglobulin that bound to antigens present in the cell culture substrate. Of marked concern was the frequent presence of cross-reactive antibodies in encephalitis cases associated with West Nile and Powassan flaviviruses. Because IFA testing for rabies on human specimens may result in false-positive results, it should not be used as the sole basis for initiating antirabies treatment.

Detection of rabies viral RNA by TaqMan real-time RT-PCR using non-neural specimens from dogs infected with rabies virus

To determine the burden of rabies in developing countries, a reliable and accurate diagnostic test for the examination of the brains of animals is needed. Recently, the number of samples and carcasses submitted to rabies diagnostic units has been declining. Methods for obtaining tissues from different regions of the brain are even more difficult, and direct florescent antibody examination may fail if the samples decomposed. The spread of rabies virus to peripheral non-nervous tissues starts early during the pre-clinical phase. It has been shown that saliva and skin biopsies taken at the neck and containing hair follicles can be used in the ante-mortem diagnosis of rabies in humans. Obtaining oral swab samples, whisker or hair follicles from the heads of canines is easy and practical and can be performed without special equipment. The objective of this study was to determine whether these non-neural specimens can be used for the detection of rabies viral RNA. The RNAs extracted from these specimens were tested using a real-time reverse transcriptase polymerase chain reaction (RT-PCR). The sensitivity of the TaqMan real-time RT-PCR analysis using samples from dogs confirmed to be infected with rabies virus was 84.6% (55/65), 81.8% (54/66) and 66.7% (44/66) when using oral swab samples, extracted whisker follicles and extracted hair follicles; the specificity of all specimen types was 100%. The negative predictive values were 77.8%, 74.4% and 61.4%, respectively. Although the rate of positivity when combining the three non-neural specimen types was increased to 86.4%, this level of sensitivity was not sufficient to help physicians whether to administer post exposure prophylaxis. However, these oral swab and whisker specimens may serve to enhance epidemiological surveillance; such data will contribute in the planning of rabies control programs.

Rapid diagnosis of rabies in humans and animals by a dot blot enzyme immunoassay

Rapid diagnosis of rabies in humans and animals by a dot blot enzyme immunoassay

Rapid diagnosis of rabies in humans and animals by a dot blot enzyme immunoassay

The presently advocated tests for rapid diagnosis of rabies, such as the fluorescent antibody test (FAT) are expensive and require expertise to carry out and interpret the results. In this study, a simple direct dot blot enzyme immunoassay (DIA) has been developed and evaluated to detect the rabies antigen in brain specimens of animals and humans. The utility of this test in the ante-mortem diagnosis of human rabies has also been evaluated. Brain homogenates of suspected rabid animals (n = 250), humans (n = 16) and clinical samples like saliva (n = 12) and cerebrospinal fluid (CSF) (n = 12) were directly spotted on polyvinylidene difluoride membrane (PVDF) and the absorbed rabies nucleoprotein antigen was detected using biotinylated antinucleoprotein antibody followed by treatment with streptavidin peroxidase conjugate and color development with diamino benzedine (DAB). Rabies-infected and normal mouse brain homogenates were used as positive and negative controls, respectively. The results of this test were evaluated with fluorescent antibody technique (for brain samples) and mouse inoculation test (for saliva and CSF samples). A distinct dark brown color was seen in the positive control and all positive samples, while there was no color development with either the negative control or the negative samples. The concordance between the fluorescent antibody test (FAT) and dot immunoassay was 98.4% for brain samples, 83.3% for saliva and 91.6% for CSF samples. The specificity of the test was found to be 100%. The dot blot enzyme immunoassay (DIA) test described here is a sensitive, specific and rapid test for the post-mortem diagnosis of rabies in animals and humans. The utility of this test for the ante-mortem diagnosis of rabies needs to be further evaluated.

Diagnosis of human rabies by the cornea test.

The second death due to rabies encephalitis occurring among American servicemen stationed in the Republic of Vietnam is reported. The clinical diagnosis was confirmed by performing the direct fluorescent antibody test for rabies on smears of corneal epithelial cells obtained on the second day of hospitalization. The reliability of various laboratory procedures in confirming the diagnosis of human rabies early in the course of clinical illness is discussed. The cornea test is a useful and rapid method of diagnosing human rabies prior to the development of significant serum antibody titers.