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Author’s Response: Allopregnanolone Assays

University of Siena Siena 53100, Italy

To the editor:

First, we recognize that the general methodological concepts expressed by Dr. Pearson Murphy have been known for many years because they are the fundamental principles of a correct endocrinological investigation in vitro and in vivo.

In her recent letter Dr. Pearson Murphy asked how the assay for allopregnanolone is specific. The method used to measure serum allopregnanolone by Luisi et al. (1) is identical to that described by Purdy et al. (2). They used different polyclonal antiserum raised in sheep, and not in rabbit, against allopregnanolone carboxymenthyl ether coupled to BSA.

This antiserum shows a very low cross-reactivity with other steroids tested (3). Therefore, it was used also to perform other studies that have investigated healthy women (4), patients with amenorrhoea (5) or with premenstrual syndrome (6).

Our preliminary investigations suggested that to determine allopregnanolone in human serum or/and animal organs (7) is not necessary to perform preliminary purification by high-performance liquid chromatography.

Then, comparing the data obtained by Purdy et al. (2) with ours (4), it is clear that our data are significantly lower, suggesting no cross-reactivity with other steroids.

Received December 18, 2000.

References

1. Luisi S, Petraglia F, Benedetto C, et al. 2000 Serum allopregnanolone levels in pregnant women: changes during pregnancy, at delivery and in hypertensive patients. J Clin Endocrinol Metab. 85:2429–2433.[Abstract/Free Full Text]
2. Purdy RH, Moore Jr PH, Rao PN, et al. 1990 Radioimmunoassay of 3-hydroxy-5-pregnan-20-one in rat and human plasma. Steroids. 55:290–296.[CrossRef][Medline]
3. Bernardi F, Salvestroni C, Casarosa E, et al. 1998 Aging is associated with changes of allopregnanolone concentrations in brain, endocrine glands and serum in male rats. Eur J Endocrinol. 138:316–321.[Abstract]
4. Genazzani AR, Petraglia F, Bernardi F, et al. 1998 Circulating levels of allopregnanolone in humans: gender, age and endocrine influences. J Clin Endocrinol Metab. 83:2099–2103.[Abstract/Free Full Text]
5. Meczekalski B, Tonetti A, Monteleone P, et al. 1999 Hypothalamic amenorrhea with normal body weight: ACTH, allopregnanolone and cortisol responses to corticotropin-releasing hormone test. Eur J Endocrinol. 142:280–285.
6. Monteleone P, Luisi S, Tonetti A, et al. 1999 Allopregnonolone levels and premenstrual syndrome. Eur J Endocrinol. 142:269–273.
7. Genazzani AR, Bernardi F, Stomati M, et al. 2000 Effects of estradiol and raloxifene analog on brain, adrenal and serum allopregnanolone content in fertile and ovariectomized female rat. Neuroendocrinology. 72:162–170.

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