Infection of Cattle with Parainfluenza 3 Virus with Special Reference to Udder Infection

Kawakami, Yoshimi; Kaji, Tomohiro; Kume, T.; Omuro, Mamoru; Hiramune, Takashi; Murase, Nobuo; Matumoto, Minoru

doi:10.1111/j.1348-0421.1966.tb00304.x

Cited by 16 publications

(16 citation statements)

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“…An occurrence of EHV-1 abortion among imported mares in 1967 caused the first outbreak of EHV-1 abortion among pregnant mares raised in Japan [5]. Thereafter both EHV-1 and -4 spread among various horse populations in Japan.…”

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Diagnosis and Sero-Epizootiology of Equine Herpesvirus Type 1 and Type 4 Infections in Japan Using a Type-Specific ELISA.

et al. 1998

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ABSTRACT. Recently, a type-specific ELISA using equine herpesvirus type 1 (EHV-1) and type 4 (EHV-4) glycoprotein Gs (gGs) was developed by Crabb and Studdert [1993]. To investigate the dissemination of EHV-1 and -4 among horses in Japan, we applied their ELISA as suitable for discriminating between EHV-1 and -4 infections serologically. Type-specificity of the ELISA was confirmed by using paired sera of infected horses with either EHV-1 or -4. Application of the ELISA to sera collected before and after the winter season of 1995-1996 from 80 racehorses revealed that 30 horses showed significant antibody responses against EHV-1 and 9 against EHV-4, respectively. The results indicated that this ELISA using paired sera is useful for a diagnosis and an epizootiological study on EHV-1 and -4 infections. -KEY WORDS: diagnosis, epizootiology, equine herpesvirus.J. Vet. Med. Sci. 60(10): 1133-1137, 1998 Virological diagnosis can identify the type of infected virus, but they are time-consuming and laborious. Therefore, it is considered to be very important to diagnose the type of infected virus serologically. The purpose of this study was to develop a type-specific serological test for diagnosis of EHV-1 and -4 infections, which facilitates the elucidation of epizootiological characteristics of both infections among the various horse populations in Japan.The EHV-1 89c25p strain [9] and an EHV-4 strain TH20, the Japanese prototype of EHV-4 [4], were used in this study. Viruses were propagated at low multiplicities of infection in primary fetal horse kidney cells. Infected cells showing a cytopathic effect were treated with 1% sodium dodecyl sulfate (SDS) and proteinase K (0.1 mg/ml) in 0.1 M Tris-HCl (pH 8.0), 0.1M NaCl, 5 mM EDTA at 37°C overnight. Following the DNA extraction with phenol and chloroform-isoamyl alcohol (24:1), ethanol precipitated preparations were dissolved in water.Crabb and Studdert [3] reported that type-specific, continuous epitopes were located in the corresponding Cterminal variable region of EHV-1 gG (amino acids 288 to 350) and EHV-4 gG (amino acids 287 to 382), which show little amino acid identity. To express the type-specific antigen, the bacterial expression plasmid pGEX-5X1 was used. The recombinant pGEX-5X1 plasmids, pGEX-gG1P and pGEX-gG4P, were constructed according to the methods described previously [3] with a minor modification. (i) pGEX-gG1P was constructed by PCR amplification (35 cycles of 92°C for 1 min, 55°C for 1 min, 72°C for 1 min) with the EHV-1 89c25p DNA as the template and the following primers: forward primer, gG1P-F, containing an EcoRI site, 5'-CCGAATTCGGTGACGAAACATACGA-3', and reverse primer, gG1P-R, containing an XhoI site, 5'-AACTCGAGCTGGATGCCGTTCGACG-3'. The amplified fragment was digested with EcoRI and XhoI and cloned into the pGEX-5X1 digested with the same restriction endonucleases. (ii) pGEX-gG4P was constructed by PCR amplification (35 cycles of 92°C for 1 min, 50°C for 1 min,

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“…[1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16] in which a tentative serological diagnosis of EHV-1 or -4 infection was made by complement fixation (CF) test. The result showed that a final diagnosis of EHV-1 infection was made in 8 horses and that diagnosis of EHV-1 infection might be made in another 3 horses (Table 1).…”

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Diagnosis and Sero-Epizootiology of Equine Herpesvirus Type 1 and Type 4 Infections in Japan Using a Type-Specific ELISA.

et al. 1998

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“…Strain BNM-7 of parainfluenza type 3, one of the isolates from milk reported in our previous paper [6], was employed. Fluids from infected bovine kidney cell cultures at the 7th to 10th passages were used as the inoculum.…”

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confidence: 99%

“…Fluids from infected bovine kidney cell cultures at the 7th to 10th passages were used as the inoculum. Assays of virus and of neutralizing and hemagglutination-inhibiting (HI) antibodies were carried out by the methods described in the previous report [6].…”

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