Human metapneumovirus (hMPV): an associated etiology of severe acute respiratory infection in children of Eastern Uttar Pradesh, India

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Abstract

Acute respiratory infections (ARIs) are a serious public health concern across the world, causing considerable morbidity and mortality. Every year, around 13 million children under the age of five die. Approximately 95% of them are from developing nations, and ARIs are responsible for one-third of all deaths. Human Metapneumovirus (hMPV) is one of the causative agents associated with respiratory tract infections. There is lack of information about hMPV from the eastern region of Uttar Pradesh. At Indian Council of Medical Research- Regional Medical Research Centre, Gorakhpur (ICMR‐RMRC, Gorakhpur) in Uttar Pradesh, India; we tested respiratory pathogens in under-five patients presenting with ARI and severe acute respiratory illness (SARI) through semi nested PCR. A total of 100 nasal and throat specimens were collected from the outdoor and indoor patient Departments (OPD) and (IPD) of Department of Paediatrics, BRD Medical College, Gorakhpur during February to April 2022. Out of 100 enrolled paediatric patients, 4(4%) were found to be hMPV positive. Among the patients who tested positive for hMPV, 25%(1/4) unfortunately died. The phylogenetic analysis of hMPV showed the close resemblance with the clade of Singapore and USA hMPV isolates. The study underlines the importance of hMPV as the cause of acute respiratory infections in children and the highlight the need for routine testing for this virus in laboratories. Further more comprehensive detailed study on various aspects of hMPV in this area is needed.

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  1. Thank you very much for submitting your revised manuscript to Access Microbiology and promptly include the suggested changes in the text. I am pleased to let you know that the manuscript is now accepted for publication. Congratulations!

  2. Thank you very much for for submitting your revised manuscript to Access Microbiology including the amendments proposed by the reviewers, which has improved the quality of the work. However, there some minor aspects that need to be addressed. - Please adequately refer to the supplementary table (originally Table 2). As it is the only item in the supplementary files, it should be named Supplementary Table S1 and should be referred as such in the main text. - Clearly state in the appropriate section that clinical information for patients that tested negative for hMPV was not obtained from hospital. - Include in the proportion of patients presenting ARI and SARI in the main text (see see Reviewer 2, comment 5 and the author's reply to this comment). - Please consider including in the discussion section a reasoning for why 96 of the patients presented ARI or SARI but tested negative for all possible causative pathogens tested. Please provide a revised version of the manuscript (including a tracked-changes version) and a point-by-point response to these comments within 1 month.

  3. Comments to Author

    General comment The article 'Human Metapneumovirus (hMPV): An associated etiology of Severe Acute Respiratory Infection in Children of Eastern Uttar Pradesh, India.' described the detection and prevalence of Human Metapenumovirus (hPMV) in Children from Eastern Uttar Pradedh India. They used both nested PCR and sanger sequencing to detect the matrix genes of hPMV in four out of 100 patients samples tested. They showed that sequences aligned with the sequences from Singapore and the US. hPMV is an important respiratory pathogen in children and immunocompromised patients. The first detection in Utter pradedh India is important because it create awareness and should lead to increase surveillance and detection. While this report is important for global public health surveillance, I think that this article will benefit from extensive review, structural organization of the sections and grammatical review to be ready for publication. I will recommend this article for publication but with a major revision due to the following observations below. The style used for the result section is not consistent, the article started with summary style and later introduced subheadings (Line 136). The article will benefit if the author reviews the required style or structure for the journal and follows it accordingly. Secondly, I think description of patient should not be in the result it is patient metadata and such should preferably be in the methodology or supplementary section. Furthermore, the result for the respiratory diagnostic panels were not presented. It says they were all negative, so I am curious to know what the aetiology of the ARI or SARI for the 96 patients could be if they were negative for hPMV. This would have been an important result and point of discussion. In the discussion, the authors highlighted other studies that detected hPMV, however there was no detailed discussion comparing both studies. The implication of the clinical and biochemistry parameters of the positive patients versus the negative patients was not discussed. If something is highlighted in the result, then it should be important enough to be discussed. Interestingly, one of the patients died, however, there was no mention of the clinical parameters of the patient versus the other three and the 96 negative patients. Also, phyologenetic tree and the implication of the alignment of the sequences with those from the US and Singapore was elucidated. These important details would have made the discussion section ronbust. Lastly, the claim to provide new insight into the epidemiology of hPMV in India is an over interpretation. There is little or no epidemiological finding in the article. The transmission dynamics, the probable source of hPMV and other components of epidemiology relating to the pathogen was not discussed or mentioned. The key finding is the first detection of hPMV in the location. Other specific observations are below: The number of ARI versus SARI cases among the 100 patients was not mentioned. Was all the SARI cases hPMV positive or how many percentage of SARI was positive and why? Line 24 delete 'of' Line 26 positive for what? Line 47: In Chennai, 47 South India a study was conducted in children with ARI and during 2016-2018 among 350 nasal 48 swab specimens, in 4% (14/350) hMPV was detected (6). - rewrite Line 56-57 - which previous report? No reference Line 67- the reference is not the WHO definition of ARI or SARI Line 72 - please add the catalogue number of the kit that was used. Line 73-4 Please describe the semi nested PCR or give the citation of the method. Table 2- was clinical and biochemistry done for HPMV negative sample how does that compare to the one presented. Line 103- Table 1 is not a description of semi nested PCR but rather than the primers used. Line 138- it showed detection of HPMV not circulation- you didn't measure transmission to demonstrate circulation. Line 140- You need line typesetting. Line 150- Please write in past tense not past participle 'was' instead of 'has been' Line 151- mentioned a similar study with low incidence but no discussion for the difference. Line 159- grammar check (less findings...) Line 189- No new insight was provided for the epidemiology of the hPMNV

    Please rate the manuscript for methodological rigour

    Poor

    Please rate the quality of the presentation and structure of the manuscript

    Satisfactory

    To what extent are the conclusions supported by the data?

    Partially support

    Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?

    No

    Is there a potential financial or other conflict of interest between yourself and the author(s)?

    No

    If the manuscript involved human and/or animal experimentation, have the subjects been treated in an ethical manner and has ethical permission been provided?

    Yes: There is statement of ethical approval in the manuscript

  4. Thank you very much for submitting your manuscript to Access Microbiology. It has now been reviewed by two experts in the field, whose comments are attached at the bottom of this email. Overall, the reviewers consider this manuscript a valuable piece of work. However, they propose some major changes, including changes in the structure of the manuscript, improvements in the discussion and re-assessment of conclusions that may be considered overstatements. They also suggest an extensive revision of the grammar and typo correction. Please provide a revised version of the manuscript (including a tracked-changes version) and a point-by-point response to the reviewer comment within 2 months.

  5. Comments to Author

    This is a well-written article that provided the important information about Human Metapneumovirus (hMPV) in under-five patients acute respiratory infections (ARIs) with from the eastern region of Uttar Pradesh India for the first time. I recommended a minor amendment for it as following. Line 49-53, this paragraph should be supported by a reference. Line 59-61, 'We have amplified matrix protein gene of hMPV, through conventional PCR, sequenced and Phylogenetic analysis were done.' should be 'We have amplified matrix protein gene of hMPV through conventional RT-PCR, and then the sequencing and phylogenetic analysis for the product were done.' Line 73-74, 'The samples were initially screened for hMPV RNA by conventional semi nested PCR using primers (Table 1) based on matrix protein (M) gene.' The hMPV RNA should be changed to DNA by reverse transcription firstly, and then subjected to conventional semi nested PCR using primers (Table 1) based on matrix protein (M) gene. Line 75-77, Table 1 should be presented as a three-line table. The title of the table is placed at the top of the table. Line 102-103, 'All these aetiologies were found negative and further confirmed by the hMPV genus specific semi-nested PCR.' should be changed to 'All these aetiologies were found negative and the samples were further screened by the hMPV genus specific semi-nested RT-PCR.' Line 115-116, 'The patient condition improved and discharged on satisfactory grounds after 3 days of hospital admission.' should be changed to 'The patient condition improved and he was discharged on satisfactory grounds after 3 days of hospital admission.' Line 112, Patient 1, 'A 24 months male patient' to be 'A 24-month-old male patient' Similarly, Patient 2/3/4 was a 60-month-old male patient/a 5-month-old female patient/a 44-month-old female patient, respectively. Line 120, the patient condition improved and he was discharged. Line 128, Patient was given Oxygen support via NP for one day. What is NP? Line 137-139, 'The sequences show close resemblance with the clade of Singapore and USA hMPV isolates, it clearly showed the circulation of hMPV virus in pediatric population of eastern Uttar Pradesh, India.' should be 'The sequences show close resemblance with the clade of Singapore and USA hMPV isolates, clearly showing the circulation of hMPV virus in pediatric population of eastern Uttar Pradesh, India.' 'The Matrix gene sequence of the PCR amplified product used in the phylogenetic analysis.' should be 'The PCR amplified product of Matrix gene sequence used in the phylogenetic analysis.' Line 133-135, Table 2 should be presented as a three-line table. The title of the table is placed at the top of the table. Line 145, a legend should be added to Figure 1 to describe the name rules for the sample strains and reference strains. Line 149, 'hMPV has been detected globally since its discovery in 2001.' Should be supported by an appropriate reference.

    Please rate the manuscript for methodological rigour

    Good

    Please rate the quality of the presentation and structure of the manuscript

    Good

    To what extent are the conclusions supported by the data?

    Partially support

    Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?

    No

    Is there a potential financial or other conflict of interest between yourself and the author(s)?

    No

    If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?

    Yes