Immunological diagnosis of pregnancy in the mare by agglutination of latex particles

TWERIOGENOLOGY IMMUNOLOGICAL

2. 1

2

CosterI,

Cambiaso'

of

Faculty

of

45,

des

Unit

Veterinary

support,

sera

PARTICLESx

&

P.L.

Pathology

Masson'

of

Reproduction,

of

Cellular

and

Molecular

Pathology,

7430, Hippocrate,

Brussels

rue

BY

Medicine,

(Belgium)

'Acknowledgements

1,

MARE

(Belgium)

Institute

avenue

B-1200

and

THE

Medicine,

Experimental

UCL-ICP

LATEX

IN

Veterinaires,

International

75,

C.L.

Obstetrics

Brussels

of

PREGNANCY

OF

Department

B-1070

OF

AGGLUTINATION

De

rue

DIAGNOSIS

to du

and

: we Dr.

P. Delahaut

Carmel, to

correcting

Dr. the

are

to

IRSIA

(Laboratoire

Marloie) C.

grateful

for

Richards

Dr.

financial

d'Hormonologie,

collecting

and

for

and

H.W.

Holy

testing

some

for

paper.

Received for publication:

3/24/W

Abstract Pregnant in

mare

mare sera

serum

by

gonadotrophin

agglutination

anti-PMSG

immunoglobulins.

sensitive

and

rapid

correlated

perfectly

biological

test

inhibition

test.

of

This

was

applied

with

and

with

The

(PMSG)

of

test

those those was

particles

test,

which

to of of

can

latex

324 the the

positive

be

is

samples.

determined coated

with

simple, The

results

Aschheim-Zondeck hemagglutination from

the

38th

day

pregnancy.

JUNE 1980 VOL. 13 NO. 6

433

THERIOGENOLOGY INTRODUCTION Pregnancy pregnant

in

mares

mare

biological

serum

immunological

when

a

and

equipment. the

purpose

of

which

MATERIAL

work

was

be

easily

were

the

al.,

1977

O.lM bovine

434

the

a

a

method

latex

but

The

agglutination

sensitive,

PMSG

and

test.

the

Rivanol

coated

rapid.

buffer

latex 9.0

bi-monthly

(6600 in

IU/mg),

0.5

were

ml

bled

a

of

after

two

gave

(6,9-diaminoacridine precipitation-technique

(0.8 as

~1 from

described

spontaneous

the

by

rabbits,

Belgium)

avoid

pH

of

animals

particles

g/l

obtained

suspended

latex

by

were

to 100

glycine-NaOH, serum

of

special

procedure.

inoculated

Brussels,

To

mg

The

for

polystyrene

(7).

0.625

three

France)

at

preparation

attractive

was

Holland,

the

guanidinium-HCl

particles

of

isolated

Federa,

Courbevoie,

of

difficult.

delicate develop

been

of

requires

performed,

adjuvant. Of

suitable IgG

to

by

between

be

an

have

drawback

the

but

antiserum

Oss,

Freund's months.

antisera

lM

anti-PMSG

Organon,

complete

a

or

METHODS

injections

from

and

can

is

be

this

AND

lactate,

can

the

its

obvious

inhibition has

possible

would

intradermal

(6)

enzyme

rabbit

The

also

the

distinction

Enzymoimmunoassay

labeling

and

first

sedimentation is

(1,Z)

techniques

whereas

and

by

rats

have

Two

of

either

female

latter

hour-incubation,

Radioimmunoassay

determination

(PMSG)

ease.

The

erythrocytes

agglutination

by

in

(3)

(4,5). 12

PMSG-coated

three

and

: immunodiffusion

requiring

gift

The

rapidity

hemagglutination

The

detected

tested

reactions. of

proposed

be

gonadotrophin

activity

advantages

test

can

RhBne-Poulenc, by

Cambiaso

agglutination used

to

containing

resuspend This

suspension. 0.17M

we

NaCl

et added

the

buffer and

was IO

g/l

albumin.

JUNE 1980 VOL. 13 NO. 6

THERIOGENOLOGY

As

13%

were

pl of

solution

of

test

of the

pretreated min.

(1 mg/ml NaCl).

The

white

opalescent

and

particles. used

the

08512. sera

were

The

of

were

the

was

kept

AND

PMSG

batch

to that

one

group

with

those

correlation the

pregnant

83

from

between

Inc., 38 and of

7

the

non-agglutinated

inhibition

detectable

was

used

to about

test

we

Cranburg,

N.J.,

120 days

after

animal.

The

the

(4),

down

and

to

to prepare

1 IU/ml.

of biological

of

150

the

This

methods

the

163 mares

of the was

agar

ng/ml

of

antiserum

sensitivity (8),

double

inhibition

diffusion

mare,

the

later.

In a second

hemagglutination

Both

tests

were

reported

were

group

positive

JUNE 1980 VOL. 13 NO. 6

results test.

38th

test

were

in either

the

between

animals,

on the

biological

latex

Aschheim-Zondeck

observed

positive

whereas of

distinguished

after

(3).

In a first perfect

read

C.

was

which

hemagglutination

detected

was

of the

condition

25 ul

DISCUSSION

similar

compared

plate

25 ~1 of the

Carter-Wallace,

at -20"

corresponded

technique

easily aspect

of the

25 ul of a buffer

on a dark with

agglutination

collected

agglutinator,

with

glycine-NaOH

hemagglutination

from

agglutination

which was

clumps

homogeneous

irrespective

RESULTS

the

The

test

non-specific

by mixing

they

Aldrich-Europe,

2.0 min

suspension

serum.

For MIP

Blood

mating

latex

for

in O.lM

performed

mare

the

incubated

DTT

(w/v)

non-specifically, (DTT,

inactivate

was

0.17M was

10%

agglutinated

dithiothreitol to

serum

containing The

sera

with

Belgium)

Beerse, 300

of the

treated

with

day,

became

of also

both one

latex

found

in 71 cases.

A

tests

which

exception. was

positive

161 mares

were

latex

In

positive

only and

to correlate No false

16 days inhibition perfectly. negatives

group.

435

THERIOGENOLOGY

In addition non-bred

to the

mares

None

of

these

test

was

not

324

of which sera

sera

we

70 were

104

samples

at different

agglutinated

sensitive

tested

latex,

to pituitary

from

stages

suggesting

of

heat.

that

the

gonadotrophins.

REFERENCES 1. Aschheim, aus

Harn

S.,

Hormons. 2. Cole,

and

maturity

57-68

B.

des

Wschr.

7:

Hart,

Die

Schwangerschaftdiagnose

Hypophysen

1404-1411

G.

in progressive

sexual 3.

Zondeck,

Nachweis

Klin.

H.H.,

mares

and

durch

The

stages

- Vorderlappen

(1928).

potency

of blood

of pregnancy

of the

immature

rat.

Simple

immunological

serum

of

in effecting

Am.

J.

the

Physiol.

-93:

(1930).

Richards,

C.B.

diagnosis

of pregnancy

in mares.

method Nature

for

215:

the

1280-1281

(1967). 4. Allen,

W.R.

pregnant 591

A quantitative

mare

serum

immunological

gonadotrophin.

J.

assay

for

Endocrin.

-43:

581-

(1969).

5. Wide,

M.,

by an

and

Wide,

immunological

6. Schultze, of Human

H.E.,

and

Proteins.

L.

Diagnosis

method.

of pregnancy

Nature

198:

Heremans,

J.F.

Elsevier,

Amsterdam.

in mares

1017-1018

Molecular

(1963).

Biology

1966

Vol.

1

p 262. 7. Cambiaso, J. and

C.L.,

Masson,

Leek, P.L.

I. A general

method

antigens

haptens.

and

A.E.,

De Steenwinkel,

Particle for

the J.

counting

Billen,

immunoassay.

determination

Immunol.

F.,

Methods

of antibodies, -18:

33-44

(1977). 8. Vindevogel, pour

gravide.

436

H.

la mise

Comparaison

en evidence

Ann.

Med.

Vet.,

de deux

du

PMS

116:

dans 83-91

tests

biologiques

le serum

de jument

(1972).

JUNE

1980 VOL. 13 NO. 6