Serum Analysis of Hydro-pericardium Syndrome (HPS) by Filter-Paper Blood Sampling Technique: As Substitute to Serum Collection Technique

  • Abdul Razak Veterinary Research Institute, Zarrar Shaheed Road, Lahore Cantt, Punjab, Pakistan, Postal Code 54810,
  • Iftikhar Hussain 2Institute of Microbiology, University of Agriculture Faisalabad, Punjab, Pakistan, Postal Code 38000.
  • Rehan Rafique Veterinary Research Institute, Zarrar Shaheed Road, Lahore Cantt, Punjab, Pakistan, Postal Code 54810,
  • Sajid Hussain Veterinary Research Institute, Zarrar Shaheed Road, Lahore Cantt, Punjab, Pakistan, Postal Code 54810,
  • Muhammed Shoaib Noor Veterinary Research Institute, Zarrar Shaheed Road, Lahore Cantt, Punjab, Pakistan, Postal Code 54810,
  • Asaad Nayyer Veterinary Research Institute, Zarrar Shaheed Road, Lahore Cantt, Punjab, Pakistan, Postal Code 54810,
Keywords: Isolation, HPS antigen, IHA, AGPT, Filter paper sampling technique

Abstract

Background: The study aimed to assess the filter paper blood sampling technique for sero- monitoring against Hydro-pericardium syndrome (HPS). In view of the fact, dried blood samples don’t require immediate refrigeration, occupy little space and are easily transported.
Methods: Whole blood paired with serum samples were collected from 100 broiler chickens on filter paper strips, dried for 2hrs at 37oC, stored in polythene bag and then eluated in normal saline at 4oC for overnight. Antigen of HPS was isolated, purified and confirmed by agar gel precipitation test (AGPT) with raised hyper-immune serum. Eluates of whole blood dried on filter paper with corresponding serum samples were tested for antibody activity by indirect heam-agglutination (IHA) test. Results: ‘The IHA titers of eluates were similar to titers obtained with serum diluted as 1:10. Normal saline and phosphate buffered saline did not influence the antibody stability, when used as eluating fluid. Whole blood dried on filter paper could be stored sealed in plastic bag at 4oC or ambient temperature for at least one week with no appreciable loss of antibody titers. A strong correlation (r = 0.900) exist between the titers obtained with two methods of blood sampling.

Conclusion: This study demonstrated that recovery of antibodies from blood dried on filter paper after eluation produces results comparable to those obtained by recovering antibodies from serum. Based on above findings it is concluded that filter paper blood sampling could serve as a cost effective and convenient tool for HPS sero-monitoring.

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Dilutions
Negat ive

1: 2

1: 4

1: 8

1: 16

1:3 2

1:6 4

1:1 28

1:25 6

1:51 2

1:10 24

GM T

Serum samples
1
-
2
2
4
5
5
3

3

-
-


32. 89


Dilutions
Negat ive
1: 10
1: 20
1: 40
1: 80
1:1 60
1:3 20
1:6 40
1:12 80
1:25 60
1:51 20
GM T
Eluates (4°C)
4
3
4
5
3
3
2
1

-

-
-


28. 45


Eluates (ambient temperat ure)
4
3
5
5
2
2
3
1
-
-
-
27. 67

Dilutio n

Negat ive

1:2
1:4

1:8

1:1 6

1:3 2


1:6 4


1:1 28

1:25 6

1:51 2

1:10 24



GM T


Seru m sampl es
3
_
2
2
3
3
2
5
4
1
_
32. 00
Dilutio ns

Negat ive
1:1 0
1:2 0


1:4 0
1:8 0

1:1 60

1:3 20
1:6 40

1:12 80

1:25 60

1:51 20
GM T
Eluat es (1 day)
6
1
2
3
4
6
2
1
_
_
_
30. 37
Eluat es (3 days)
7
1
2

3
2
5

3
2
_
_
_


28. 47

Eluat es (6 days)

7
1


2


4

2


5

2

2


_


_

_
26. 20
Statistically no effect of storage time on stability of antibodies
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International Archives of BioMedical And Clinical Research
Vol 5 | Issue 1 | January – March 2019 52

www.iabcr.org
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Published
2019-03-21
How to Cite
1.
Razak A, Hussain I, Rafique R, Hussain S, Noor MS, Nayyer A. Serum Analysis of Hydro-pericardium Syndrome (HPS) by Filter-Paper Blood Sampling Technique: As Substitute to Serum Collection Technique. Int Arch BioMed Clin Res [Internet]. 2019Mar.21 [cited 2019Oct.17];5(1):50-3. Available from: https://iabcr.org/index.php/iabcr/article/view/466
Section
ORIGINAL ARTICLES ~ Allied Health Sciences