scholarly journals Relation Between the Polymerase Chain Reaction and the Indirect Fluorescent Antibody Method in the Diagnosis of Legionella Infection

1996 ◽  
Vol 23 (3) ◽  
pp. 656-657 ◽  
Author(s):  
M. Koide ◽  
A. Saito ◽  
N. Kusano ◽  
M. Tateyama ◽  
J. Inadome ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Fluorescent Antibody ◽  
Indirect Fluorescent Antibody ◽  
Chain Reaction ◽  
Antibody Method ◽  
Polymerase Chain ◽  
Fluorescent Antibody Method

scholarly journals Persistence of Porcine Reproductive and Respiratory Syndrome Virus in Serum and Semen of Adult Boars

1995 ◽  
Vol 7 (4) ◽  
pp. 456-464 ◽  
Author(s):  
Jane Christopher-Hennings ◽  
Eric A. Nelson ◽  
Rebecca J. Hines ◽  
Julie K. Nelson ◽  
Sabrina L. Swenson ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Virus Isolation ◽  
Fluorescent Antibody ◽  
Acute Stage ◽  
Viral Rna ◽  
Respiratory Syndrome Virus ◽  
Indirect Fluorescent Antibody ◽  
Chain Reaction ◽  
Serum Samples ◽  
Polymerase Chain

Four seronegative adult boars were intranasally inoculated with porcine reproductive and respiratory syndrome virus (PRRSV) isolate VR-2332. Serum and semen were collected 2-3 times weekly for over 100 days postinoculation (DPI). Serum samples were assayed for PRRSV by virus isolation (VI) and a polymerase chain reaction (PCR) and screened for antibodies to PRRSV using the indirect fluorescent antibody (IFA) and virus neutralization (VN) tests. Semen was assayed for PRRSV RNA by PCR. Virus or viral RNA was detected in the serum of all boars within 1 DPI by VI and/or PCR. However, VI results indicated that viremia was transient and occurred from 1 to 9 DPI. Viral RNA was detected in serum from 1 to 31 DPI. In the acute stage of the infection, PRRSV RNA was detected in serum by PCR prior to the presence of viral RNA in semen. The PRRSV RNA was detected in semen as early as 3 DPI and persisted for 25 DPI in 2 of the boars and 56 and 92 DPI in the remaining 2 boars. Detection of PRRSV RNA in semen occurred 2-8 and 28-35 days prior to the detection of antibodies by IFA and VN, respectively. PRRSV was isolated from the bulbourethral gland of the boar that shed viral RNA in semen for 92 DPI. These results suggest that PRRSV RNA can be detected by PCR in boar serum and semen, and may persist for variable periods of time. Viremia and the serologic status of the boar are not adequate indicators of when PRRSV or PRRSV RNA is being shed in the semen. Preliminary findings also indicated that neither shipping stress nor reinoculation with homologous PRRSV resulted in viremia or viral RNA shedding in semen.

Comparison of 10 Indirect Fluorescent Antibodies to Detect and Type Influenza A Specimens

2010 ◽  
Vol 134 (8) ◽  
pp. 1177-1180
Author(s):  
Rosemary C. She ◽  
Edward W. Taggart ◽  
Cathy A. Petti
Keyword(s):  
Polymerase Chain Reaction ◽  
Disease Control ◽  
Reverse Transcription ◽  
Influenza A ◽  
Fluorescent Antibody ◽  
Indirect Fluorescent Antibody ◽  
Chain Reaction ◽  
Specific Antibodies ◽  
Control And Prevention ◽  
Polymerase Chain

Abstract Context.—Management of influenza infections relies on rapid, accurate, and sensitive diagnostic techniques. Influenza A (IA) strain typing has become more important since the emergence of highly pathogenic avian and novel influenza strains and the high frequency of oseltamivir resistance in circulating H1N1 isolates. Objective.—To analyze the performance of indirect fluorescent antibody testing for subtyping a broad range of IA strains. Design.—Ten indirect fluorescent antibody reagents were used to detect and type 100 archived IA respiratory specimens from 1986 through 1995 and 2006 through 2007 and a reassortant, nonpathogenic H5N1 sample. Both direct specimen and cultured isolates were tested. Reverse transcription-polymerase chain reaction was used to confirm indirect fluorescent antibody results. Three H1N1-, 2 H3N2-, and 1 H1-H2-H3-H5–specific antibodies (Chemicon Diagnostics), an IA pool reagent (Trinity Biotech), and H1, H3, and H1-H3–specific antibodies (Centers for Disease Control and Prevention) were used. Results.—Reverse transcription-polymerase chain reaction confirmed all 100 isolates as IA and identified 71 as H1, 22 as H3, and 7 as non–H1-H3. Sensitivity of direct specimen testing ranged was 18.3% to 57.7% for the H1 reagents, 36.4% to 50.0% for the H3 reagents, and 40.0% to 53.8% for the pool reagents. Subtyping was more sensitive on cultured isolates than direct specimens. Specificity for all antibodies was 89.7% to 100%. The H5N1 sample was positive by direct testing and culture (reverse transcription-polymerase chain reaction, Centers for Disease Control and Prevention H5N1 pool, Chemicon H1-H2-H3-H5). No cross-reactivity was observed when the 10 antibodies were tested against other common respiratory viruses. Conclusions.—When positive, IA subtyping antibodies can serve as a useful diagnostic tool when multiple influenza virus subtypes are cocirculating with different susceptibility patterns.

scholarly journals A Comparative Analysis of Polymerase Chain Reaction and Direct Fluorescent Antibody Test for Diagnosis of Genital Herpes

2017 ◽  
Vol 9 (01) ◽  
pp. 053-056 ◽  
Author(s):  
Vrushali Patwardhan ◽  
Preena Bhalla ◽  
Deepti Rawat ◽  
Vijay Kumar Garg ◽  
Kabir Sardana ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Genital Herpes ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Conventional Polymerase Chain Reaction ◽  
Genital Ulcer ◽  
Chain Reaction ◽  
Direct Fluorescent Antibody ◽  
Polymerase Chain ◽  
Simplex Virus

ABSTRACT Objective: To compare laboratory tests that can simultaneously detect and type herpes simplex virus (HSV) directly from the genital ulcer specimens in clinically suspected cases of genital herpes. Materials and Methods: A study was conducted over 10 months and 44 adult male and female patients clinically suspected with genital herpes were recruited. Genital ulcer swab specimens were subjected to glycoprotein-G gene-based conventional polymerase chain reaction (PCR) and commercially available direct fluorescent antibody (DFA) test and the results were compared. Results: PCR for HSV was positive in 82% (36/44) cases. DFA was positive in 68.2% (30/44) cases. There was 100% agreement between HSV types detected by DFA and PCR. The strength of agreement between the results was better in primary genital herpes than recurrent cases. Conclusion: PCR was found to be better in the detection of HSV in recurrent genital herpes patients. It is a better modality, especially when genital herpes clinically presents with ulcerative or crusted lesions, and is also a cheaper alternative as compared to DFA.

scholarly journals Cattle rabies: the effect of clinical evolution, viral genetic lineage, and viral load on the severity of histological lesions

2020 ◽  
Vol 40 (4) ◽  
pp. 227-233
Author(s):  
Claudia S. Wisser ◽  
André Thaler Neto ◽  
Helena B.C.R. Batista ◽  
Enio Mori ◽  
Maria E.R. Chierato ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Fluorescent Antibody ◽  
Inflammatory Lesion ◽  
Genetic Lineage ◽  
Fluorescent Antibody Technique ◽  
Chain Reaction ◽  
Antibody Technique ◽  
Qrt Pcr ◽  
Polymerase Chain

ABSTRACT: Our objective was the characterization and staging of histological lesions in different anatomical sites of the central nervous system (CNS) of rabid cattle. The severity of the lesions was compared with the clinical stages of the disease, the variants of viral isolates, and with the load of virus. Thirty-one spontaneously affected rabid cattle the state of Santa Catarina underwent clinical follow-up and were eventually necropsied. CNS tissues were sampled and submitted to direct fluorescent antibody technique (DFAT), immunohistochemistry (IHC), routine histopathology with hematoxylin and eosin stain (HE), reverse transcriptase polymerase chain reaction (RT-PCR), and polymerase chain reaction in quantitative reverse transcriptase in real time (qRT-PCR). Affected cattle were allotted in four groups according to their clinical stage when euthanized: G1, euthanized while standing; G2, euthanized when in sternal recumbence; G3, euthanized when in lateral recumbence; and G4, affected cattle with natural death. In order to evaluate the degree of severity of the lesions and the presence of Negri bodies (NBs), the brain was sectioned at 9 sites. Additionally, spinal cord and trigeminal ganglion sections were examined. The intensity of the lesions was graded as either absent, mild, moderate, or marked, and the presence or absence of the NBs was noted. Histological lesions were characterized by lymphocytic and monocytic meningoencephalitis with NBs in 28 cases. In all analyzed groups, intensities of histological lesions ranging from mild to severe were observed. Brain regions with the highest inflammatory lesion intensity were the medulla at the level of obex, followed by the colliculus and thalamus. NBs were observed in a higher percentage in the cerebellum, followed by medulla at the obex level, striatum complex, and frontal telencephalon. The duration of the clinical course of the disease did not influence the intensity of the inflammatory lesion, but it did influence the presence of NBs, with a higher percentage of these inclusions in cattle that died naturally than in euthanized cattle. All isolated rhabdovirus included in this study were genetically compatible with samples from hematophagous bats Desmodus rotundus. The evaluation by qRT-PCR did not demonstrate a correlation between lesion intensity and the amount of virus.

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