Monastir le 25/11/2008 - Infectious Agents and Cancer

Author's response to reviews
Title: Quantitative analysis of human herpesvirus-6 genome in blood and bone
marrow samples from Tunisian patients with acute leukemia: A Follow-up Study
Authors:
Faten Nefzi ([email protected])
Agnès Gautheret ([email protected])
Nadia Ben Fredj ([email protected])
Nabil Abid Ben Salem ([email protected])
Monia Zaier ([email protected])
Abderrahim Khelif ([email protected])
Agut Henri ([email protected])
Salma Feki ([email protected])
Mahjoub Aouni ([email protected])
Version: 2 Date: 9 October 2012
Author's response to reviews: see over
République Tunisienne
Université de Monastir
*****
*****
Ministère de l’Enseignement Supérieur
Faculté de Pharmacie
de la Recherche Scientifique et de la
*****
Laboratoire des Maladies
Technologie
*****
Secrétariat d’état de la Recherche
Directeur : Pr. Mahjoub
Scientifique et de la Technologie
AOUNI
Transmissibles & Substances
Biologiquement Actives
Cover letter
Professor Franco M Buonaguro
Edito-in-Chef
Infectious Agents and Cancer
Tunisia, 09th October 2012
Dear Professor,
We thank the reviewers for making corrections and suggestions that will improve our
manuscript.We haverevised it, carefully, according to all your comments and suggestions.We have
a great pleasure to resubmit it. The corrections as well as the additions were underlined in the
current revised version.
Answers to the questions from the first reviewer (Sylvie Hermouet)
Comment 1
The paper would benefit from editing and improving of the English. It would also benefit from a
more concise format. For example, HHV-6 copy numbers and % of patients found positive for
HHV-6 DNA are cited several times in the manuscript, notably in Discussion (page 11, lines 6-3)
where they should be deleted.
Response 1
The manuscript has been reviewed by a native English speaker.
The manuscript has been modified to a more concise format, in particular in Abstract, Results and
Discussions sections.
Comment 2
The authors should state how well the qPCR techniques perform in their hands. For instance readers
expect to know the sensitivity of qPCRs, ie. the lowest number of copies detected. Indeed,
Laboratoire des Maladies Transmissibles et Substances Biologiquement Actives, Faculté de Pharmacie, Rue Avicenne 5000 Monastir, Tunisie
Tél. : + 21 6 73 461 000 / + 216 73 466 950 Fax : + 216 73 465 754 E-Mail :[email protected]
République Tunisienne
Université de Monastir
*****
*****
Ministère de l’Enseignement Supérieur
Faculté de Pharmacie
de la Recherche Scientifique et de la
*****
Laboratoire des Maladies
Technologie
*****
Secrétariat d’état de la Recherche
Directeur : Pr. Mahjoub
Scientifique et de la Technologie
AOUNI
Transmissibles & Substances
Biologiquement Actives
besides the variability in patient cohorts, different qPCR sensitivities can explain that some studies
found 36% leukemic patients positive for HHV-6 DNA whereas the present study finds only 15%
(Discussion, page 11).
Response 2
The quantitative PCR and the PCR for type identification have been performed in the laboratory of
virology from Pitié-Salpêtrière hospital, where the methods had been developed. The sensitivity of
detection and quantitation are those usually obtained in the laboratory (10 copies of HHV-6
genome). We totally agree with the reviewer concerning the explanation for the differences of
sensitivity between the study of Hermouet et al and our study. This point has been added in the
discussion (page 11, line 194–195).
Comment 3
Similarly, Nefzi et al. report that 100% of their HHV-6-positive patients carried the B genotype,
which differs from other published studies. Again, this difference might be due to the patient cohort,
but the authors must provide evidence that the published technique performed in their lab can
indeed detect HHV-6A, and they should provide the lowest number of HHV-6A copies detected.
Response3
Since the sensitivity of the qPCR used is identical for HHV-6B and HHV-6A, identification of only
HHV-6B may be linked to the high number of children included in this study, and to the high
prevalence for these patients. This point has been added in the discussion (page 12, line 226–230).
The lowest number of HHV-6A copies detected was 10 copies of HHV-6A genome (page 16, line
290-291).
Comment 4
The authors should better describe the case with CI-HHV-6: provide biological and clinical
information, at diagnosis, response to treatment, etc…; they should provide cytogenetics data. It
Laboratoire des Maladies Transmissibles et Substances Biologiquement Actives, Faculté de Pharmacie, Rue Avicenne 5000 Monastir, Tunisie
Tél. : + 21 6 73 461 000 / + 216 73 466 950 Fax : + 216 73 465 754 E-Mail :[email protected]
République Tunisienne
Université de Monastir
*****
*****
Ministère de l’Enseignement Supérieur
Faculté de Pharmacie
de la Recherche Scientifique et de la
*****
Laboratoire des Maladies
Technologie
*****
Secrétariat d’état de la Recherche
Directeur : Pr. Mahjoub
Scientifique et de la Technologie
AOUNI
Transmissibles & Substances
Biologiquement Actives
would also be very interesting to document transmission of HHV-6 DNA in other members of the
patient’s family.
Response 4
Complement data concerning the patient with CI-HHV-6 have been added in the result section
(page 9, 10 line 175–182).
Comment 5
Tables 2 & 3 could be merged in a single table.
Response 5
Tables 2 and 3 have been merged in a single table (page 21).
Comment 6
Discussion, page 12, line 19: “genotype” is missing (“HHV-6 B was the only …identified…”).
Response 6
The sentence has been reformulated (page 12, line 226-230).
Answers to the questions from the second reviewer (Maria LinaTornesello)
Comment 1
The writing style in the whole manuscript must be more concise. For example in the abstract the
three sentences (lines 11-13) “In blood samples, HHV-6 was detected in 15% of patients at
diagnosis. At aplasia, 8% of ….” can be just condensed in one sentence: “In blood samples, HHV-6
was detected in 15% of patients at diagnosis, in 8% during aplasia, in 30 % at remission and in 28%
at relapse”. Please revise the whole manuscript and modify the text accordingly.
Laboratoire des Maladies Transmissibles et Substances Biologiquement Actives, Faculté de Pharmacie, Rue Avicenne 5000 Monastir, Tunisie
Tél. : + 21 6 73 461 000 / + 216 73 466 950 Fax : + 216 73 465 754 E-Mail :[email protected]
République Tunisienne
Université de Monastir
*****
*****
Ministère de l’Enseignement Supérieur
Faculté de Pharmacie
de la Recherche Scientifique et de la
*****
Laboratoire des Maladies
Technologie
*****
Secrétariat d’état de la Recherche
Directeur : Pr. Mahjoub
Scientifique et de la Technologie
AOUNI
Transmissibles & Substances
Biologiquement Actives
Response 1
The manuscript has been modified to a more concise format, in particular in Abstract, Results and
Discussions sections.
Comment 2
Abstract. Line 13 -15 The abbreviation used for copies per million cells is atypical. The standard
abbreviation is “copies/million cell". Text should be modified as follow “Median viral load was 138
copies/million cells at diagnosis, 244 at aplasia, 112 at remission and 78 copies/million cells at
relapse”.
Response 2
The abbreviation used for HHV-6 viral load has been modified from "Cop/M" to “copies/million
cells".
Comment 3
Abstract. Line 16. To avoid continuous repetitions the sentence could be rephrased as follow:
“Median viral loads were 34, 109 and 32 copies/million cells at diagnosis, remission and relapse,
respectively.”
Response 3
The sentence was rephrased as follow “The median viral loads were 34, 109 and 32 copies/million
cells at diagnosis, remission and relapse, respectively” (page 3, line 51–52).
Comment 4
Abstract. Line 17.%19??may be 19%
Response 4
In the abstract, "%19" has been changed for "19%" (page 3, line 53).
Comment 5
Abstract. Line 19. The words "with median viral loads at 206 Cop/M" and "with median viral loads
at 79 Cop/M" should be put in parenthesis to make the sentence more clear.
Laboratoire des Maladies Transmissibles et Substances Biologiquement Actives, Faculté de Pharmacie, Rue Avicenne 5000 Monastir, Tunisie
Tél. : + 21 6 73 461 000 / + 216 73 466 950 Fax : + 216 73 465 754 E-Mail :[email protected]
République Tunisienne
Université de Monastir
*****
*****
Ministère de l’Enseignement Supérieur
Faculté de Pharmacie
de la Recherche Scientifique et de la
*****
Laboratoire des Maladies
Technologie
*****
Secrétariat d’état de la Recherche
Directeur : Pr. Mahjoub
Scientifique et de la Technologie
AOUNI
Transmissibles & Substances
Biologiquement Actives
Response 5
In the abstract, the words "with median viral loads at 206 Cop/M" and "with median viral loads at
79 Cop/M" have been put in parenthesis (page 3, line 53-54).
Comment 6
Abstract. Line 21. The sentence “HHV-6 was more detected . ..” should be changed in “HHV-6 was
more prevalent in …”.
Response 6
In the abstract, the sentence “HHV-6 was more detected ...” has been changed for “HHV-6 was
more prevalent in …” (page 4, line 57).
Comment 7
Abstract. Line 23. Clarify the sentence “HHV-6 detected in patients was HHV-6B”. Do the authors
mean all patients? Specify.
Response 7
In the abstract, the sentence “HHV-6 detected in patients was HHV-6B” has been changed to be
more precise (page 4, line 58).
Comment 8
Introduction pg 6 line 7. Indicate which values of “Median viral loads were found in patients with
lymphoproliferative”.
Response 8
In the introduction the values of “Median viral loads were found in patients with
lymphoproliferative” have been added (page 6, line 97, 98).
Comment 9
Results pg 7. All the section “Clinical samples” must be moved to the Methods section.
Laboratoire des Maladies Transmissibles et Substances Biologiquement Actives, Faculté de Pharmacie, Rue Avicenne 5000 Monastir, Tunisie
Tél. : + 21 6 73 461 000 / + 216 73 466 950 Fax : + 216 73 465 754 E-Mail :[email protected]
République Tunisienne
Université de Monastir
*****
*****
Ministère de l’Enseignement Supérieur
Faculté de Pharmacie
de la Recherche Scientifique et de la
*****
Laboratoire des Maladies
Technologie
*****
Secrétariat d’état de la Recherche
Directeur : Pr. Mahjoub
Scientifique et de la Technologie
AOUNI
Transmissibles & Substances
Biologiquement Actives
Response 9
In the previous Results section, the section “Clinical samples” has been moved to the Methods
section (page 14, line 248-255, 260-264).
Comment 10
Discussion pg 11. Repetition of data already described in the Results section should be mostly
removed.
Response 10
The discussion section has been modified to avoid repetition of data already described in the
Results section.
Quality of written English: Needs some language corrections before being published
The manuscript has been reviewed by a native English speaker.
Please, find as attached files our revised manuscript
Sincerely Yours,
Faten NEFZI
Laboratoire des Maladies Transmissibles et Substances Biologiquement Actives, Faculté de Pharmacie, Rue Avicenne 5000 Monastir, Tunisie
Tél. : + 21 6 73 461 000 / + 216 73 466 950 Fax : + 216 73 465 754 E-Mail :[email protected]