J Neurol Res

Journal of Neurology Research, ISSN 1923-2845 print, 1923-2853 online, Open Access

Article copyright, the authors; Journal compilation copyright, J Neurol Res and Elmer Press Inc

Journal website http://www.neurores.org

Original Article

Volume 2, Number 1, February 2012, pages 16-24

The Possible Underworld of Chronic Fatigue Syndrome From Neurotransmitters Polymorphisms to Disease

**Table 1.** Primer Pairs and Restriction Enzyme Used for Genetic Analysis
Polymorphism	Primer Sequence	Annealing	Amplified Fragment Size (bp)	Restriction Enzyme
SLC6A4 5HTLPR	GGCGTTGCCGCTCTGAATGC GAGGGACTGAGCTGGACAACCAC	61 °C	484 (S allele) 528 (L allele) 613 (XL allele)
HTR2A -1438 G/A	AAGCTGCAACCTAGCAACAGC AACCAACTTATTTCCTACCAC	60 °C	468	HpaII
BDNF Val66Met	AGAAGAGGAGGCTCCAAAGG CTGATTTGTGTCTGGTGCAG	60 °C	499	PmlI
GSK3B -50 T/C	CTCGCTTCCTTCCTTCCTTT GATTCCCAGACGCCTGTTAC	58 °C	237	AluI
GSK3B -157 T/C	CAGCTGCTTTGCACTAACAGA TTAGGTGACAAACGCTTTCTTT	55 °C	250	Hpy188I

Table 1. Primer Pairs and Restriction Enzyme Used for Genetic Analysis

Polymorphism

Primer Sequence

Annealing

Amplified Fragment Size (bp)

Restriction Enzyme

SLC6A4 5HTLPR

GGCGTTGCCGCTCTGAATGC
GAGGGACTGAGCTGGACAACCAC

61 °C

484 (S allele)
528 (L allele)
613 (XL allele)

HTR2A -1438 G/A

AAGCTGCAACCTAGCAACAGC
AACCAACTTATTTCCTACCAC

60 °C

468

HpaII

BDNF Val66Met

AGAAGAGGAGGCTCCAAAGG
CTGATTTGTGTCTGGTGCAG

60 °C

499

PmlI

GSK3B -50 T/C

CTCGCTTCCTTCCTTCCTTT
GATTCCCAGACGCCTGTTAC

58 °C

237

AluI

GSK3B -157 T/C

CAGCTGCTTTGCACTAACAGA
TTAGGTGACAAACGCTTTCTTT

55 °C

250

Hpy188I

**Table 2.** Clinical Characteristics and Presence of XMRV of 70 Italian CFS Patients
Clinical Features	% (n = 70)
Age of onset	33.5 ± 12.5
Male	23%
Female	77%
Arthralgia	89%
Myalgia	8%
Depressive symptoms	43%
Unexplained fever	60%
Comorbidities
Type 2 Diabetes	3%
Hypothyroidisms	4%
Rheumatoid Arthritis	1%
Psoriasis	1%

Table 2. Clinical Characteristics and Presence of XMRV of 70 Italian CFS Patients

Clinical Features

% (n = 70)

Age of onset

33.5 ± 12.5

Male

23%

Female

77%

Arthralgia

89%

Myalgia

Depressive symptoms

43%

Unexplained fever

60%

Comorbidities

Type 2 Diabetes

Hypothyroidisms

Rheumatoid Arthritis

Psoriasis

**Table 3.** Allelic and Genotypic Distribution Between CFS Patients and Controls
	CFS patients (n = 70)	Controls (n = 128)	P value
SLC6A4 5-HTTLPR
LL	34%	29%	ns
LS	47%	53%	ns
SS	18%	18%	ns
L	58%	55%	ns
S	42%	45%	ns
HTR2A -1438 G/A
GG	23%	17%	ns
GA	48%	56%	ns
AA	29%	17%	ns
G	47%	50%	ns
A	53%	50%	ns
BDNF Val66Met
GG	50%	51%	ns
GA	34%	32%	ns
AA	11%	17%	ns
G	64%	67%	ns
A	36%	33%	ns
GSK3B -50 T/C
TT	46%	39%	ns
TC	44%	40%	ns
CC	10%	15%	ns
T	68%	63%	ns
C	32%	37%	ns
GSK3B -157 T/C
TT	63%	46%	ns
TC	27%	39%	ns
CC	10%	15%	ns
T	76%	66%	ns
C	24%	34%	ns

Table 3. Allelic and Genotypic Distribution Between CFS Patients and Controls

CFS patients (n = 70)

Controls (n = 128)

P value

SLC6A4 5-HTTLPR

34%

29%

47%

53%

18%

58%

55%

42%

45%

HTR2A -1438 G/A

23%

17%

48%

56%

29%

17%

47%

50%

53%

50%

BDNF Val66Met

50%

51%

34%

32%

11%

17%

64%

67%

36%

33%

GSK3B -50 T/C

46%

39%

44%

40%

10%

15%

68%

63%

32%

37%

GSK3B -157 T/C

63%

46%

27%

39%

10%

15%

76%

66%

24%

34%

**Table 4.** Allelic and Genotypic Distribution Between Male and Female CFS Patients
		Male N = 20	Female N = 44	χ²	P value
SLC6A4	LL	50%	27%	2.2215	0.136
5-HTTLPR	LS	30%	57%	2.9586	0.085
	SS	20%	16%	0.002	0.964
	L	65%	56%	0.6376	0.425
	S	35%	44%
HTR2A	GG	30%	18%	0.5386	0.463
-1438G/A	GA	40%	52%	0.4106	0.522
	AA	30%	30%	0.0667	0.796
	G	50%	44%	0.1652	0.684
	A	50%	56%
BDNF	GG	60%	52%	0.0929	0.761
VAL66MET	GA	25%	41%	0.8996	0.343
	AA	15%	7%	0.3344	0.563
	G	73%	73%	0.035	0.852
	A	28%	27%
GSK3B	TT	60%	45%	0.6545	0.419
-50TC	TC	30%	45%	0.1576	0.691
	CC	10%	9%	0.0918	0.762
	T	75%	68%	0.3294	0.566
	C	25%	32%
GSK3B	TT	35%	59%	2.3034	0.129
-157TC	TC	55%	30%	2.7927	0.095
	CC	10%	11%	0.0729	0.787
	T	63%	74%	1.2004	0.273
	C	38%	26%

Table 4. Allelic and Genotypic Distribution Between Male and Female CFS Patients

Male
N = 20

Female
N = 44

χ²

P value

SLC6A4

50%

27%

2.2215

0.136

5-HTTLPR

30%

57%

2.9586

0.085

20%

16%

0.002

0.964

65%

56%

0.6376

0.425

35%

44%

HTR2A

30%

18%

0.5386

0.463

-1438G/A

40%

52%

0.4106

0.522

30%

0.0667

0.796

50%

44%

0.1652

0.684

50%

56%

BDNF

60%

52%

0.0929

0.761

VAL66MET

25%

41%

0.8996

0.343

15%

0.3344

0.563

73%

0.035

0.852

28%

27%

GSK3B

60%

45%

0.6545

0.419

-50TC

30%

45%

0.1576

0.691

10%

0.0918

0.762

75%

68%

0.3294

0.566

25%

32%

GSK3B

35%

59%

2.3034

0.129

-157TC

55%

30%

2.7927

0.095

10%

11%

0.0729

0.787

63%

74%

1.2004

0.273

38%

26%

**Table 5.** Allelic and Genotypic Distribution Between CFS Patients With and Without Depressive Symptoms
	CFS patients with depressive symptoms	CFS patients without depressive symptoms	CHI	P	OR	95% CI
	male	12%	65%	14.3023	0.0002	0.074	0.0184 - 0.3005
	female	88%	35%			13.444	3.3274 - 54.3234
5-HTTLPR	LL	36%	26%	0.2495	0.62
	LS	48%	53%	0.0125	0.91
	SS	16%	21%	0.0119	0.91
	L	60%	53%	0.3313	0.56
	S	40%	47%
HTR2A	GG	16%	21%	0.0119	0.91
-1438G/A	GA	36%	59%	2.159	0.14
	AA	48%	21%	3.7822	0.05	3.56	1.1351 - 11.1675
	G	34%	50%	2.3891	0.12
	A	66%	50%
BDNF	GG	52%	62%	0.2337	0.63
VAL66MET	GA	40%	32%	0.1096	0.74
	AA	8%	6%	0.0417	0.84
	G	72%	78%	0.275	0.60
	A	28%	22%
GSK3B	TT	40%	44%	0.0025	0.96
-50TC	TC	52%	44%	0.1125	0.74
	CC	8%	12%	0.0002	0.99
	T	66%	66%	0.0312	0.86
	C	34%	34%
GSK3B	TT	68%	56%	0.4529	0.50
-157TC	TC	28%	29%	0.0298	0.86
	CC	4%	15%	0.8256	0.36
	T	82%	71%	1.4555	0.23
	C	18%	29%

Table 5. Allelic and Genotypic Distribution Between CFS Patients With and Without Depressive Symptoms

CFS patients with depressive symptoms

CFS patients without depressive symptoms

CHI

95% CI

male

12%

65%

14.3023

0.0002

0.074

0.0184 - 0.3005

female

88%

35%

13.444

3.3274 - 54.3234

5-HTTLPR

36%

26%

0.2495

0.62

48%

53%

0.0125

0.91

16%

21%

0.0119

0.91

60%

53%

0.3313

0.56

40%

47%

HTR2A

16%

21%

0.0119

0.91

-1438G/A

36%

59%

2.159

0.14

48%

21%

3.7822

0.05

3.56

1.1351 - 11.1675

34%

50%

2.3891

0.12

66%

50%

BDNF

52%

62%

0.2337

0.63

VAL66MET

40%

32%

0.1096

0.74

0.0417

0.84

72%

78%

0.275

0.60

28%

22%

GSK3B

40%

44%

0.0025

0.96

-50TC

52%

44%

0.1125

0.74

12%

0.0002

0.99

66%

0.0312

0.86

34%

GSK3B

68%

56%

0.4529

0.50

-157TC

28%

29%

0.0298

0.86

15%

0.8256

0.36

82%

71%

1.4555

0.23

18%

29%